JPH0543697B2 - - Google Patents

Abstract

Polypeptide derivatives containing 5-amino-2,5-disubstituted-4-hydroxypentanoic acid residues are of the formula:wherein, for example in a preferred embodiment R is t-butyloxycarbonyl, m and n are each 0 or 1, their sum being at least 1; R is cyclohexylmethyl; R² is 2-methylpropyl, 2-methyl-2--propenyl, benzyl, 4-chlorobenzyl, 3-chlorobenzyl, 4-methylbenzyl, 4-methoxybenzyl, 3,4-dichlorobenzyl, 2-chlorobenzyl or cyclohexylmethyl; R³ is hydrogen; R⁴ is hydrogen, methyl, 4-aminobutyl, 2-carboxyethyl, 3-(ethoxycarbonyl)propyl or 3-(benzyloxycarbonyl)propyl; W is phenylalanyl, W¹ is histidyl and W² is lysyl.These compounds are useful for inhibiting the angiotensinogen cleaving action of the enzyme renin having particular utility in the treatment of hypertension.

Description

[Detailed description of the invention]

INDUSTRIAL APPLICATION The present invention describes a series of novel polypeptide derivatives containing 5-amino-2,5-disubstituted-4-hydroxyvaleric acid residues useful for inhibiting the angiotensinogen cleavage activity of the enzyme renin. This invention relates to a method for stereoselective production of intermediates. Prior Art The proteolytic enzyme renin (molecular weight approximately 40,000) is produced in the kidneys and secreted into the blood. This substance is known to have the activity of cleaving angiotensinogen, a natural plasma glycoprotein, in vivo. In the case of human angiotensinogen, it is cleaved at the bond between leucine (10th position) and valine (11th position) at the N-terminus of angiotensinogen: Asp 1- Arg 2- Val 3- Tyr 4 - Ile 5- His 6- Pro 7- Phe 8- His 9- Leu 10- Val 11- Ile 12- His 13- Ser 14- Glu 15- N-terminal decapeptide formed by the above cleavage action by renin ( Angiotensin) circulates and is broken down in the body into an octapeptide known as angiotensin. Angiotensin is known as a potent pressor substance (i.e., a substance that can cause a significant increase in blood pressure) and is thought to act by causing constriction of blood vessels and secretion of the sodium-retaining hormone aldosterone from the adrenal glands. . The renin-angiotensinogen system has therefore been implicated as a contributing factor in certain forms of hypertension and congestive heart failure. One means of reducing the detrimental effects of the renin-angiotensinogen system is the administration of substrates that can inhibit the angiotensinogen cleavage action of renin. Many such substrates are known, including anti-renin antibodies, pepstatin, and natural phospholipid compounds. European Patent Application No. 77028 describes non-terminal statin (Sta; 4-amino-3-hydroxy-
A series of renin-inhibiting polypeptide compounds are disclosed, including statin derivatives (6-methylvaleric acid) or statin derivatives. The majority of the compounds exemplified, including Sta, have six or more amino acid residues. A few examples of short chains disclosed therein include Acetyl-Phe-His-Sta-Leu-Phe-NH ₂
and t-butyroxycarbonyl-Phe-His-
Examples include Sta-Leu-Phe- _NH2 . There are always at least two amino acid residues on either side of the statin. Statyl groups in di- or polypeptides are always attached to fat-soluble amino acids (mostly leucine). (See also U.S. Pat. Nos. 4,470,971 and 4,478,826.) European Patent Application No. 45,665 and U.S. Pat.
No. 4424207 is the formula (wherein A is e.g. t-butoxycarbonyl, B is His or other basic aminoacyl group, D is Val, Ile or other lipophilic aminoacyl residue, E is Tyr, Phe, His or other aromatic aminoacyl residues, where R ^a and R ^b are each isopropyl, isobutyl, benzyl or other lipophilic amino acid type side chains, and Y ^a is a terminal acid, ester or amide type group). A series of renin inhibitory polypeptide derivatives are disclosed. Including the central 5-aminovaleric acid residue, these compounds are invariably heptapeptides, i.e., N
-tetrapeptidyl-5-aminopentanoyl fat-soluble aminoacyl-aromatic amino acid derivative. A Solution to the Problem We have now discovered that a class of new compounds has special value as renin inhibitors. These compounds have the formula or a pharmaceutically suitable salt thereof [wherein n and m are each 1 or 0, and the sum of n and m is at least 1; W is , where R ⁵ is phenyl, 1-naphthyl,
4-hydroxyphenyl, 4-methoxyphenyl, 4-imidazolyl, propyl or isopropyl; W ¹ is

[expression] or and W ² is and R ⁶ is −NH ₂ ,

[Formula] or -CH ₂ NH ₂ ; When n is 1, R is hydrogen, an amino-protected acyl group with a molecular weight of less than 500, prolyl, pyroglutamyl, or the nitrogen protected by the above-mentioned amino-protected acyl group. prolyl or pyroglutamyl; when n is 0, R is phenoxyacetyl or 2-benzyl-3-phenylpropionyl (dibenzylacetyl); R ¹ and R ² are independently hydrogen, (C ₁ -C ₆ )alkyl, (C ₁ -C ₆ )alkenyl, phenyl, naphthyl, (C ₄ -C ₇ )cycloalkyl, (C ₄ -C ₇ )cycloalkenyl, (C ₇ -C ₉ ) enyl alkyl,
( _C11 - _C13 ) naphthylalkyl, ( _C5 - _C10 )(cycloalkyl)alkyl, ( _C5 - _C10 )(cycloalkenyl)alkyl, or ( _C1 - _C3 )alkyl,
(C ₁ -C ₃ ) One of the above groups in which the aromatic ring is mono- or di-substituted by the same or different groups selected from alkoxy, fluoro or chloro; (a) R ³ and R ⁴ are , separately hydrogen, ( _C1 - _C6 ) alkyl, phenyl, naphthyl, ( _C4 - _C7 )cycloalkyl, ( _C7 - _C9 ) phenylalkyl,
( _C11 - _C13 )naphthylalkyl, ( _C5 - _C10 )(cycloalkyl)alkyl or adamantyl, or ^R3 is hydrogen and ^R4 is [Here, p and q are independently 0 or 1 to
is an integer of 6; r is 0 or 1; Q is _-CH2- , -CH=CH-, -O-, -NH
-, -CHOH- or -CO-; Y is methyl, phenyl, -COOR ⁹ , -CONR ⁹
R ¹⁰ , −CONHCOOCH ₂ C ₆ H ₅ , NH ₂ , −
NHCOCH ₂ CH ₂ C ₆ H ₅ ,

[expression] or

[Formula]; R ⁷ , R ⁸ , R ⁹ and R ¹⁰ are independently hydrogen, (C _1
-C6 ) alkyl, phenyl, ( _C4 - _C7 ) cycloalkyl, ( _C7 - _C9 ) phenylalkyl, ( _C5)
-C ₁₀ ) (cycloalkyl)alkyl or adamantyl] or (b) R ³ or R ⁴ together with the nitrogen to which they are attached are pyrrole, indoline, isoindoline, piperidine, 1,2, 3,4-tetrahydroquinoline, 1,2,3,4-tetrahydroisoquinoline, perhydroazepine or morpholine ring system]. The expression "amino-protected acyl group" means an acyl group that can substantially inhibit the reaction at the α-nitrogen position of W (or W ¹ when n=0) in vivo. To prevent undue deleterious effects on solubility properties, R has a molecular weight of 500 or less. Examples of suitable amino-protected acyl groups include t-butyloxycarbonyl, t-butylacetyl, benzylcarbonyl, t-butylureido, (tris-hydroxy)-(t-butylureido) and phenoxyacetyl groups. Examples include those that are well known to those involved. When n is 1, a preferred R is t-butoxycarbonyl, and preferred W and W ¹ are phenylalanyl and histidyl, respectively. When m is 1, a preferred W ² is lysyl. In all cases, preferred R ¹ is cyclohexylmethyl and preferred R ² are 2-methylpropyl, benzyl, 2-methylpropenyl, 4-chlorobenzyl, 4-methylbenzyl, 4-methoxybenzyl, 3,4 -dichlorobenzyl, 2-chlorobenzyl, 3-chlorobenzyl and cyclohexylmethyl, particularly preferred is 2-methylpropyl. When m is 0 and n is 1, the preferred
NR ³ R ⁴ is NH ₂ , NHCH ₃ , N(CH ₃ ) ₂ , NHCH ₂
C ₆ H ₅ , NH (CH ₂ ) ₃ COOH, NH (CH ₂ ) ₄ , NH
_{( CH2} ) _{4NHCOCH2C6H5 and tetrahydroisoquinoline} . When m and n are each 1, preferred NR ³ R ⁴ are phenylalanine and statin ethyl ester. The expression "pharmaceutically acceptable salts"
_HCl , _HNO3 , _H2SO4 , _{H3PO4 , CH3SO3H ,
pCH3C6H4SO3H , CH3COOH or HOOCCH2 _}
Refers to an acid addition salt of a compound of formula () having a basic function in an acid such as, but not limited to, CH ₂ COOH. When compound () contains one or more basic functions, its salt optionally contains one or more equivalents of acid. In addition, when the compound of formula () contains an acidic function, the expression is an alkali metal salt (e.g., Na, K), an alkaline earth metal salt (e.g.,
It also refers to cationic salts such as, but not limited to, amine salts (e.g., diethanolamine, meglumine) or amine salts (e.g., diethanolamine, meglumine). A pharmaceutical composition comprising an effective amount of a compound of formula () as an essential active ingredient in a pharmaceutically suitable carrier and an effective amount of a compound of formula () for inhibiting renin in a mammal by administering the effective amount of a compound of formula () to a mammal. It can inhibit the cleavage of angiotensinogen. As an intermediate for producing compounds of formula () [where R ¹ and R ² are defined above; W ³ is Here, R ¹⁴ is −NHCO ₂ CH ₂ CH ₂ C ₆ H ₅ ,

or -CH ₂ NHCO ₂ C ₆ H ₅ ; R ¹¹ is hydrogen or t-butoxycarbonyl; (a) R ¹² and R ¹³ are independently hydrogen, (C ₁ -C ₆ ); Alkyl, phenyl, naphthyl, ( _C4 - _C7 ) cycloalkyl, adamantyl, ( _C7 - _C9 ) phenylalkyl, ( _C11 - _C13 ) naphthylalkyl,
or (C ₅ -C ₁₀ )(cycloalkyl)alkyl, or R ¹² is hydrogen and R ¹³ is (where p, q, r and Q are defined above; Y ¹ is methyl, phenyl, -COOR ¹⁸ , -
CONR ⁹ R ¹⁰ , −CONHCOOCH ₂ C ₆ H ₅ , −
_{NHCOCH2C6H5 , _}

【formula】 or

[Formula] and; R⁷,R⁸,R⁹and R^Tenis separately defined above.
It is something that was given; R¹⁸is R other than hydrogen⁷) is the basis of; or (b)R¹²and R¹³is the nitrogen they are adding
together with pyrrole, indoline, isoy
ndrine, piperidine, 1,2,3,4-tet
Lahydroquinoline, 1,2,3,4-tetrahydrogen
Droisoquinoline, perhydroazepine or
which forms the morpholine ring system]
compound and formula There are intermediate compounds. The present invention relates to a method comprising the following stereoselective steps:
do: formula [R in the formula¹⁶(C₁−C₆) alkyl, (C₁−C₆)
alkenyl, phenyl, naphthyl, (C_Four−C₇)
Chloalkyl, (C_Four−C₇) cycloalkenyl,
(C₇−C₉) Phenylalkyl, (C₁₁−C₁₃) Nafuchi
Lualkyl, (C_Five−C_Ten)(cycloalkyl)al
Kill, or (C₁−C₃) alkyl, (C₁−C₃)a
selected from lucoxy, fluoro or chloro
one or two groups on the aromatic ring by the same or different groups.
one of the above substituted groups]
LiC≡CCOOR¹⁵[R in the formula¹⁵(C₁−C₃) alkyl
] in an inert solvent at −50° to −80°C.
React, mainly formula A process characterized by forming a compound of To obtain the compound of formula (), the following steps are further carried out:
is necessary. where X is chloro, bromo or other nucleophilic
is a substitutable group, R¹and R²is defined above
However, R²is something other than hydrogen,
Activated allyl or benzylic type groups are preferred
is; and produces pure, conventionally unavailable
The following equations (D), (E), (F), (G) and (H)
It is a chiral intermediate. A preferred X is bromo
Ru. Compounds of formula () are generally well known to those skilled in the art.
It is synthesized by the following method. Salt suitable for chemical synthesis
Protected amino acid residues of basic subunits
No α-amino group activated for acylation
It has a carboxyl group and its own α-nitrogen
Acylation with an amino acid attached to an appropriate protecting group
and there is a peptide bond between those two amino acid residues.
and then remove the above-mentioned protecting groups. this
The molecular structure of the main synthesis of subunits by binding and removal
starting from the C-terminus of the structure and repeating to the N-terminus.
and build up the polypeptide. Compounds of the invention
The standard α-amino acid used in the synthesis of products is α
- both amino-protected and α-amino unprotected forms
Also commercially available (as free acid, salt or ester, etc.)
has been done. Statins are N-(t-butyroxica)
commercially available as statin;
Statins are widely used in the literature [e.g., U.S. Pat. No. 4,397,786 and
Journal by Rich, D.H. et al.
Organ Chemistry (J.Org.Chem), No.
43, pp. 3624 (1978)].
Both γ-amino protected and γ-amino unprotected
Synthesized (as free acid or ester)
It can be done. If necessary, 4-aminobutyric acid, 4-
Aminovaleric acid, 5-(benzyloxycarbonyl)
amino)pentylamine or 3-(2-amino)
Suitable N such as ethyl ethoxy)propionate
- unprotected amino acid analogs (free acids, salts or
(e.g.) due to the reaction during the first coupling process.
Used as a chemical substance. If it is not commercially available
If the compound is not known or not known, the compound is
easily synthesized by known methods. life of purpose
The product is a basic amino acid residue W²(i.e. m=1)
If it contains, the required intermediate H-(W³)−NR¹²R¹³(W in the formula³,R¹²and R¹³
is defined above) is α−NH₂The base is
Anhydrous acidic conditions (e.g. about 3-5NHCl/dioxa
or trifluoroacetic anhydride).
such as t-butoxycarbonyl (t-Boc)
Protected by a hydrolyzable group, the ω-nitrogen is
Hydrogenation such as carbonyloxycarbonyl (cbz)
Lysine, ornitho protected by a group that can be decomposed
Amine or arginine HNR¹²R¹³and katsupu
Ring and combine. Suitable coupling conditions
is approximately an equimolar amount of the reagent to be bound, an equimolar amount of
dicyclohexylcarbodiimide (or similar
dehydrative coupling agent) and 1 to 2 molar equivalents of
1-Hydroxybenzotriazole at 0-50℃
(conveniently at room temperature) CH₂Cl₂reaction inert like
Carry out in a solvent. When the reactants are in the form of acid salts,
tertiary atom in an amount sufficient to neutralize the acid.
(e.g., triethylamine, N-methylmorpho)
phosphorus). Before the next coupling process
, basic amino acid residue W²The α-amino group of
Be protected. For example, α-t-Boc group is −10°~40
3.5-4.5NHCl/diox at °C (conveniently room temperature)
Sun effect or -20° to 20°C (conveniently 0
~5°C) by the action of trifluoroacetic anhydride.
easily removed. Essential 2,5-disubstituted-4-hydroxyvaleric acid residue
The group generally must be synthesized. main body
The selective pathway has the appropriate chirality in the first step.
(L-amino acids are convenient)
Start with: The starting material, a protected L-amino acid, is
N-(t
-Butoxycarbonyl)phenylalanine
including reduction of aromatic rings if necessary, such as
can be obtained by well-known and common methods.
Lower alkyl esters (preferred are those shown)
methyl ester), for example in toluene, from −50 to
Aqueous with isobutylaluminum hydride at −80℃
Easily reduced to rudehyde. Next, the aldehyde
LiC≡CCOOR¹⁵(Usually R¹⁵is ethyl propiolate
(Ethyl derived from
React in a reaction-inert solvent such as trahydrofuran.
make them respond. Generally, in this process, a pair of diastema
rheomer is generated, but the desired diastereomer (C)
is a big priority. A small amount of undesired isomer is
Palladium catalyst (Pb/
BaSO_Fourstandard catalytic reduction, such as
[carried out under conditions] after catalytic reduction of the triple bond
and (by reaction in toluene in the presence of acetic acid)
Preferably, chthons are formed. Next, 4 as shown in (E) and (F)Shas stereochemistry
2 to 2.5 molar equivalents of the desired lactone epimer
LiN〔CH(CH₃)₂]₂or lithium hexamethyl
in a substantial excess, such as disilazide (preferred).
In the presence of a strong base of low nucleophilicity, halide R²X
(X=Cl, Br or I; preferred is X=Br)
Matched. Halides are double bonds or aromatic rings.
Ant-shaped or bent-shaped halogen containing
compounds (e.g., 2-methyl-2-propenyl bromide)
benzyl bromide) is preferred, and if saturated
R of²If necessary, perform catalytic reduction later as shown below.
do; Again, transformer (2R) with a stereochemistry of
Preference is given to diastereomers. (Objective R²base
Before or after the catalytic reduction necessary to produce
) Separate by chromatography. Furthermore, especially
R in the condensation step²is benzyl re
The catalytic reduction completes the total synthesis of the desired compound.
It can be postponed until For catalytic reduction of simple olefins
The preferred catalyst is Pd or Rh,
Rh is preferred for reduction to chlorohexyl. In a preferred embodiment of the invention, especially NR¹²
R¹³is NH₂,NHCH₃,N(CH₃) or NHCH₂
C₆H_FiveEspecially when it is a simple amine such as
A lactone can be directly treated with its appropriate amine as follows:
react; The lactone is reacted in an inert solvent at a low temperature (e.g. 0
Excess amount of NH at ~40℃)₃, N.H.₂CH₃and N.H.
(CH₃)₂Lower molecular weight, more basic amines such as
and respond smoothly. more disturbed or weakly basic
With amines, the reaction takes place at high temperatures such as 80-100℃.
reaction is carried out, optionally in the presence of an acetic acid catalyst.
be called. More complex NR¹²R¹³And in the protected tract
(J) was acidified with dilute NaOH in an aqueous solvent.
reaction-inert substances such as dimethylformamide.
in a neutral solvent, K₂C.O.₃benzyl bromide in the presence of
Convert the acid to the benzyl ester using
Pyridinium in a reactive inert solventp-1-Sylate
3,4-dihydro-2H in the presence of a catalyst such as
- Tetrahydropyranine is produced by the action of pyran.
and finally acid It is preferable to convert the lactone by hydrogenolysis to
It is. The acid was then prepared as described above (dicyclohexyl
Dehydrative coupling (with carbodiimide)
law, or also well-known as exemplified below.
Amine compounds using the so-called mixed acid anhydride method H-(W³)_n−NR¹²R¹³ and then heated in an acetic acid aqueous solution to form a tetra
Selectively hydrolyzes hydropyranyl ether,
R¹¹of the formula () where is t-butoxycarbonyl
Synthesize intermediate compounds. Regardless of its source, R¹¹is t-Boc
selectively (by the method described above)
When cleaved to R¹¹Free amino compounds where is hydrogen
Object () was obtained. In the final step of the preferred method, this intermediate is R-W^Four−(W^Five)_o-OH (wherein R and n are as defined above;
W^Fourand W^Fiveare W and W above respectively¹to
corresponding) and (but with t-butoxycarbonyl)
Imidazole nitrogen in the protected histidine residue
) to combine. The preferred coupling method is as described above.
using dicyclohexylcarbodiimide using
use The preferred method is that when n is 1, each
than the usual method of introducing amino acids one by one.
is also particularly advantageous. Finally, imidazole t-Boc
Protecting groups are removed with aqueous acetic acid (e.g., 80% acetic acid) at 10-40°C.
selectively removed by hydrolysis in N
-benzyloxycarbonyl or benzyl es
The ter protecting group is removed by standard catalytic reduction methods as described above.
selectively removes the desired N-terminal
Leave the guard as is. All structural designations of stereochemistry shown herein
represents absolute stereochemistry. The invention is illustrated in the following examples.
For example: However, the present invention is limited to these embodiments.
Please understand that you are not limited to the details.
Must be. All temperatures are in °C, otherwise specified
If not, it is at room temperature. All solvents are removed in vacuum
This will be carried out below. All standard solutions are not otherwise specified.
If not, it's water. THF is tetrahydrofuran
stands for; DMF stands for dimethylformamide.
;DME stands for 1,2-dimethoxyethane
; DMSO represents dimethyl sulfoxide.
All standard α-amino acids or their derivatives are in the L form
It is. Statins are 3S,4S-4-amino-3-
Hydroxy-6-methylheptanoic acid. Reference example 1S -3-cyclohexyl-2-(t-butoxy
carbonylamino)-propionaldehyde S-3-cyclohexyl-2-(t-butoxy
Methyl carbonylamino)-propionate (51.2
g, 0.179 mol) was dissolved in 782 ml of dry toluene.
and cooled to -78°. Keep it at -70° or -78°.
and hydrogenated diisobutylaluminum for 1 hour.
(449 ml of 1M toluene solution, 0.449 mol)
added. Add methanol (13 ml) at −70°, followed by
and add 608 ml of half-saturated sodium potassium tartrate.
The mixture was then warmed to ambient temperature. ether
(300 ml), separate the organic layer, and saturate 1.
Washed with sodium potassium tartrate. original water
Extract the sexual layer with 600ml of fresh ether and add 600ml
backwash with fresh saturated sodium potassium tartrate of
Ta. Combine the organic layers with MgSO_FourDry on top and leave
Strip the title product,¹Based on H−nmr
Rubber obtained as a base and contaminated with toluene
45.6g; tlc Rf 0.45 (1:3 ethyl acetate;
Kisan);¹H−nmr (CDCl₃) Delta: 9.9 or
2.3(m), which is 1.4 and contains a t-butyl singlet, 3.0
−4.8(m), 4.9−5.2(d), 9.6(s). Example 1 4R.S.,5S-6-cyclohexyl-5-(t-
butoxycarbonylamino)-4-hydroxy
-2-Ethyl hexinate Dry, freshly distilled THF (117 moles)
and diisopropylamine (22.0ml, 15.8g,
0.156 mol), N₂Reaction flask dried under fire
The resulting solution was cooled to -30°,
butyl lithium (1.6M hexane solution) over 5 minutes.
76.9 ml, 0.123 mol) was added. Next, add this solution to
-78° while maintaining the temperature between -65° and -78°.
ethyl propiolate (12.5 ml,
12.1 g, 0.123 mol) was added dropwise. -78° for 30 minutes
After that, the title product of Reference Example 1 in 35 ml of THF
(19.52 g, 0.0866 mol) again from −65° to −78°
It was added over 20 minutes while maintaining the temperature. 2 hours later
Add 200 ml of 5:1 THF:acetic acid to the reaction mixture of
Then warm it up to ambient temperature and reduce the air to half capacity.
and an equal volume of 10% citric acid. Separate the organic layer and add fresh 10% citric acid sequentially.
2 x 200ml, 200ml brine and saturated NaHCO₃
2 x 200 ml, washed with MgSO_FourDry on top and dissipate
This gave 38.2 g of dark red oil. The latter,
10cm x 42cm silica gel color while doing TLC test
Chromatograph on top of 5 parts 1:9 vinegar.
Eluted with ethyl acid:hexane. Column with 1500ml
After development, fractions of 500 ml were collected. minutes
When pictures 29-37 are combined and diffused, the title product is obtained.
Obtained as an oil, 15.3 g; tlc Rf 0.44 (3:7
ethyl acetate; hexane);¹H−nmr (CDCl₃) del
Contains a singlet for the t-butyl group at 1.5
1.0-2.0 (m, 25H), 3.8-5 (m, 6H). Reference example 2 4S,5S-6-cyclohexyl-5-(t-
butyloxycarbonylamino)-gamma-he
xanolactone Title product of Example 1 (18.28g) and 5%
Pb/BaSO_Four(10.97g) with 150ml of ethyl acetate
The mixture was hydrogenated for 2 hours under 4 atmospheres of hydrogen. touch
The medium is removed by filtration, the filtrate is diffused and
interbody 4R.S.,5S-6-cyclohexyl-4-h
Droxy-5-(t-butyloxycarbonyla)
19 g of ethyl mino)hexanoate was obtained. the latter
Dissolved in 250 ml of 2.5% acetic acid in toluene,
Reflux for 3 hours, dissipate, and remove the residue.
10cm x 30cm silica gel while inspecting by TLC
Chromatographed on column with 9:11
ether:hexane 4, 1:1 ether:hexane
8, 11:9 ether:hexane 2 and finally
Elute with 3:2 ether:hexane, 28
×400ml fraction, 6×150ml fraction and finally 11×
A 400 ml fraction was collected. Combine fractions 17−24 and
When co-dissipated (costrip) with the ether, the main component
Desired, relatively low polarity 4S,5S−
The title product was obtained as an oil, 9.13 g; tlc Rf
0.5 (7:3 etherhexane), the one with greater polarity
4 of the title product ofR,5S-Epimers are combined
isolated by diffusing fraction 28−45.
and crystallized by trituration with hexane.
1.77 g; melting point 101.5-103.5°. Reference example 3 2R,4S,5S- and 2S,4S,5S−
6-cyclohexyl-5-(t-butoxycal
(bonylamino)-2-(2-methyl-2-prope
nil)-gamma-hexanolactone Dry, freshly distilled THF (30 ml) and
diisopropylamine (3.51ml, 2.52g,
0.0249 mol), N₂Reaction film dried under fire
Charge the flask and cool the resulting solution to -50°.
Butyl lithium (13.9 ml of 1.6M hexane solution,
0.0222 mol) and the mixture was further heated to −78°.
It was cooled down. Title product of Reference Example 2 in 15ml of THF
(2.77 g, 0.0089 mol) was added dropwise over 10 minutes.
and form the enolate for an additional 20 min at −78°.
3-bromine in 5 ml of THF.
-2-Methyl-1-propene was added over 10 minutes.
Ta. Stir the mixture for an additional hour at −78°;
5ml saturated NH₂Stop the reaction with Cl and bring to room temperature.
Warm and dilute with half volume of ether, 2 x 50ml
of 10% citric acid, 2 x 50 ml saturated NaHCO₃Oyo
Wash with 1 x 25 ml of brine and add MgSO._Fourdry on top
When dried and dissipated, a mixture of the title epimers
3.06g of certain oil was obtained. This latter is 7cm x 20
Chromatographed on silica gel in cm; tlc
Tested with; sequentially 1:9 ether:hexane 2,
3:17 ether:hexane 4, 1:4 ether
Le: hexane 2, 1:3 ether: hexane 2
, 7:13 ether:hexane 2 and 1:1
Elute with ether:hexane 2; 125 ml each
Fractions were collected and separated by; Trance
(2R) less polar title with stereochemistry
The compounds were collected in fractions 30-48 and these were combined.
This is obtained as an oil by dissipating it.
1.17g; tlc Rf 0.45 (2:3 ether:hexyl)
Sun);¹H−nmr (CDCl₃) Delta 1.4 (s, 9H),
1.8 (s, 3H), 0.3-3.0 (m, 18H), 3.6-4.0 (m,
1H), 4.69 (s, 1H), 4.75 (s, 1H), 4.1−4.8
(m, 2H). From fractions 55-76, the more polar
The title product is obtained, also as an oil,
0.358g, cis(2S) had stereochemistry;
tlc Rf 0.36 (2:3 ether:hexane);¹H-
nmr is the same as that of the less polar epimer.
It was hot. Reference example 4 2R,4S,5S- and 2S,4S,5S−
6-cyclohexyl-5-(t-butoxycal
Bonyl-amino)-2-(2-methylpropyl)
-gamma-hexanolactone The less polar title product of Reference Example 3 (1.17
g) and 10% Pb/C (0.351 g) in 20 ml of acetic acid.
Hydrogenate the mixture in ethyl at 4 atm for 2.5 hours,
Recover the catalyst by filtration and dissipate the filtrate
and the less polar title product (also
i.e. 2Rhas stereochemistry) is an oil
This substance crystallizes when left alone.
1.20g; melting point 88-93°; tlc Rf 0.65 (1:1
ether:hexane), Rf 0.73 (2:1 acetic ether),
Le: hexane). Sith (2S) have stereochemistry
Other isomers were obtained similarly; tlc Rf 0.59 (1:
1 ether:hexane). This Rf 0.65 (1:1
using a less polar epimer of ether (hexane).
In the following reference example that uses 2RStereochemistry is specified
Ru. Reference example 5 2R,4S,5S-6-cyclohexyl-5-
(t-butoxycarbonylamino)-4-hydro
xy-2-(2-methylpropyl)hexanoic acid
benzil 2 of the smaller polarity in Reference Example 4Rtitle product
(1.0 g, 2.72 mmol), DME (4 ml) and
Combine NaOH (4.4 ml of 5N, 0.022 mol)
After stirring for 12 hours, the organic solvent was allowed to evaporate.
Dilute the aqueous residue with water (5 ml) and add an equal volume of
Cool to 20° and cool with 22 ml of 1N HCl.
Upon acidification, the aqueous layer became cloudy by this time.
Separate this ether layer to obtain the aqueous layer (PH2.0).
Washed with an equal volume of fresh ether. ether layer
Combine MgSO_FourWhen dried on top and dissipated,
Intermediate 2R,4S,5S-6-cyclohexyl
-5-(t-butoxycarbonyl-amino)-4-
Hydroxy-2-(2-methylpropyl)hexa
0.909g (2.36g) of phosphoric acid was obtained as a white foam.
mmol). The latter, K₂C.O.₃(0.330g, 2,36 mm
0°
It was cooled to Benzyl bromide (0.308ml, 0.443g,
2.59 mmol) and stirred the mixture.
Warm up to room temperature. After 4 hours, this reaction mixture
Dilute with an equal volume of ether, 1 x 75 ml of 10%
Citric acid, 1 x 75 ml saturated NaHCO₃Wash with
MgSO_FourDry on top, dissipate, oil, 1.35 g,
I got it. The latter was placed on a 4 cm x 20 cm silica gel plate.
Romatographed; examined by TLC; 1:
9 ether:hexane 1, 3:17 ether:hexane
xane 1, 1:4 ether:hexane 1
and finally eluted with 3:7 ether:hexane 1
and fractions of 23 ml were collected. Fraction 46−75
When combined and dissipated, the crystals are suitable for recirculation.
0.583 g of solid starting material was obtained. Fraction 85−
119 and dissipate, the title product becomes an oil.
0.275g; tlc Rf 0.47 (1:1
ether: hexane);¹H−nmr (CDCl₃): 0.3−2.0
(m, 24H), 1.4 (s, 9H), 2-2.4 (m, 1H),
2.6-3.0 (m, 1H), 3.2-3.7 (m, 2H), 4.65 (d,
1H), 5.1 (s, 2H), 7.3 (s, 5H). Reference example 6 2R,4S,5S-6-cyclohexyl-5-
(t-butoxycarbonylamino)-2-(2-
methylpropyl)-4-(2-tetrahydropyra
Benzyl (oxy)hexanoate Product of Reference Example 5 (0.442 g, 0.929 mmol),
3,4-dihydro-2H-pyran (97%, 0.255
ml, 0.235 g, 2.79 mmol) and p-toluene
Pyridinium phosphate (23 mg, 0.093 mmol),
6.5ml CH₂Cl₂In a flask with a stopper.
After stirring for 18 hours at
Dissolve the distillate in 100 ml of ether and add 2 x 100 ml
Wash with half-saturated brine of MgSO_Fourdry on
Se, CH₂Cl₂Co-diffusion using
Drying gave the title product as an oil, 0.565
g; tlc Rf 0.59 (2:3 ether:hexane). Reference example 7 2R,4S,5S-6-cyclohexyl-5-
(t-butoxycarbonylamino)-2-(2-
methylpropyl)-4-(2-tetrahydropyra
Nyloxy)hexanoic acid Product of Reference Example 6 (0.565 g) and 10% Pd/
C (0.170 g) were combined in 10 ml of acetic ether.
The mixture was hydrogenated for 2.5 hours at 4 atm. Catalyst for filtration
Collect the filtrate by dipping into CH₂Cl₂co-dissipated using
The title product was obtained as an oil, 0.55 g.
This contains the theoretical yield (0.474g) of product.
;tlc Rf 0.0 (2:3 ether:hexa
), 0.35 (1:1 ethyl acetate:hexane);¹H−
nmr (CDCl₃) is 9.5−10.1(bs, 1H, COOH)
Contains peaks due to benzyl groups. This reference example was repeated on a scale of 300mg and the product
was dried under high vacuum to yield 250.5 mg
(Quantitative yield of title product as a white foam). Reference example 8 N-[N-alpha-(t-butoxycarboni
)-N-epsilon-benzyloxy-cal
Bonyl) Lysyl Phenylalanine Benzyl
Stell N-Hydroxybenzotriazole (162mg,
1.2 mmol), N-methylmorpholine (101.2 mg,
1 mmol), L-phenylalanine benzyl
Stel p-toluenesulfonate (428 mg, 1 ml)
limole), N-alpha-(t-butyloxycal)
(bonyl)-N-epsilon-benzyloxycarl
Bonyl-L-lysine (456 mg, 1.2 mmol) and
and 1-cyclohexane-3-(2-morpholinoe)
(chill) carbodiimidometh-p-toluenesulfo
salt (635 mg, 80% purity, 1.2 mmol) at 0
Sequentially dissolved in methylene chloride (50 ml) at °C to obtain
The solution was stirred at 20°C for 19 hours. This anti
The reaction mixture was then diluted with 75 ml of 5.5% HCl aqueous solution, saturated
NaHCO₃Wash with 75ml aqueous solution and 75ml brine
and MgSO_FourDry on the top, dissipate, and prepare the title raw material.
The product was obtained as a dry foam, 0.669 g;¹H−nmr
(CDCl₃) is delta 1.5 (s, 9H, t-butyl)
include. This product can be purified without further purification.
Used directly in the following reference example. Reference example 9 N-[N-epsilon-(benzyloxycarbo
lysyl] phenylalanine benzyl S
tel hydrochloride The product of Reference Example 8 (0.650g) was added to dioxane.
Dissolve in 7ml of 3.7N HCl and leave for 1 hour.
Trapping gave the title product, 0.583 g;¹H−
nmr (CD₃OD) is delta 5.2 (s, 2H, −CH₂C₆
H_Five)including. Reference example 10 N-((N-alpha-[2R,4S,5S-6
-cyclohexyl-5-(t-butoxycarbo
nylamino)-4-hydroxy-2-(2-methy
lupropyl)hexanoyl]-N-epsilon
-(benzyloxy-carbonyl)lysyl))-
Phenylalanine benzyl ester hydrochloride Product of Reference Example 9 (0.560 g, 1.01 mmol)
CH₂Cl₂Stir with 5 ml and remove under nitrogen.
Cooled to ℃. Triethylamine (0.211ml,
1.51 mmol), and then CH₂Cl₂Ginseng in 5ml
The product of Example 7 (0.474 g; 1.01 mmol) was added.
I got it. Finally, 1-hydroxybenzotriazole
(0.204 g; 1.51 mmol) and then CH₂Cl₂5
Dicyclohexylcarbodiimide (0.208 in ml)
g, 1.01 mmol) was added. Surrounding the reaction mixture
It was heated to temperature and stirred for 18 hours. like this
The resulting suspension is filtered and the filtrate is CH₂Cl₂of
Sprinkle, dilute with 15 ml of ethyl acetate and refilter.
Ta. Add the second filtrate to 2 x 10 ml of 10% citric acid, 2 x 10
ml saturated NaHCO₃and 1 x 10ml brine
Wash and MgSO_FourDry on CH₂Cl₂Using
Upon co-emission, the intermediate title product tetra
Hydro-2-pyranyl ether, 0.97g, was obtained.
Ta. The latter was stirred for 20 hours with 35 ml of 70% acetic acid.
I did. Dissipate the mixture to a low volume and remove the residue.
is co-diffused with toluene to form a solid white residue.
A residue was produced. Add this latter to 20ml of CH₂Cl₂dissolved in
1 x 10 ml brine and 2 x 10 ml saturated
NaHCO₃Wash with MgSO_FourDry on top and dissipate
This gave 1.02 g of a white solid. This is 7cm
Chromatographed on ×15 cm silica gel
CHCl₃Elute with 0.5% methanol 2 in tlc.
The samples were tested and fractions of 125 ml were collected. title product
The combined fractions 29−40 were diffused and the residue was evaporated.
It is recovered as a white solid by trituration with
0.497 g; melting point 157-159°; tlc Rf 0.39
(1:1 ethyl acetate; hexane;¹H−nmr
(CDCl₃) is 1.4(s,9H; t-butyl) and
Contains 6.5-7.4 (m, 15H, aromatic). Reference example 11 N-Alpha-((2R,4S,5S-6-shiku
lohexyl-5-[(N-t-butoxycarbonyl
ruphenyl-alanyl)histidyl]amino-
4-hydroxy-2-(2-methylpropyl)
hexanoyl))-N-epsilon-((benzyl
oxycarbonyl))lysyl-phenylalani
benzyl ester The product of Reference Example 10 (250 mg, 0.282 mmol)
Stir with 3 ml of trifluoroacetic acid,
CaCl₂Protected from moisture by drying tube and cooled to 0°
did. After 35 min, co-dissipate the reaction with ether.
and intermediate N-((N-alpha-[2R,4S，
5S-6-cyclohexyl-5-amino-4-hyron
Droxy-2-(2-methylpropyl)hexano
]-N-epsilon-(benzyloxycarbo
lysyl)) phenylalanine benzyl S
tertrifluoroacetate and obtained as a white foam,
0.291g. The latter, N₂2ml of CH at the bottom₂Cl₂dissolved in
Cool to 0° and add triethylamine (0.0471
ml, 0.0342g, 0.338mmol), 1-hydroxy
-Benzotriazole (0.0571g, 0.423 mmol)
), N-alpha-[N-(t-butoxycarboxyl),
Nyl)-phenylalanyl]-N(imidazole)-
(t-butoxycarbonyl)histidine (149.7
mg, 0.296 mmol) and CH₂Cl₂Jishi in 5ml
Chlorhexylcarbodiimide (61.1mg, 0.296mg)
mol) were added sequentially. Bring this reaction mixture to room temperature.
Heat up to 18 hours and stir for 18 hours.
The solvent of the cloudy solution was evaporated and the residue was dissolved in 5 ml of ethyl acetate.
Suspend in dicyclohexyl urea, filter and
After removing 373 mg of solids was obtained.
This results in a significant amount of hydrogen on the gamma-histidine nitrogen.
Stidineimidazole-(t-butoxy-carbo
desired intermediate product retaining the protecting group
TLC has been determined by TLC to also contain
Rf 0.51 (1:19 methanol:CHCl₃). Release the filtrate.
When scattered, tlc and¹H−nmr (CDCl₃)(two
two at 1.3 and 1.6 for the t-butyl group of
singlet and benzyl group for methylene
5.1 (indicating the singlet line in
A residue containing 140 mg was obtained. between both
Combine the substances contained in the body and stir under nitrogen.
et CHCl₃Treat with 70ml of saturated dimethylamine for 3 hours.
I understood. This reaction mixture is then diffused to
A solid with no odor was obtained and 7 cm x 15 cm of silica gel was obtained.
Chromatograph above and CHCl₃2 or two of them
Gradient elution with 5% methanol and check by TLC.
Ta. The product fractions were combined, diffused and diluted with hexane/
Trituration with ether gave the title product as a white solid.
0.182g; tlc Rf 0.85 (1:1:1:1 vinegar
Ethyl acid: Butanol: Acetic acid: H₂O), Rf 0.24
(1:19 methanol:CHCl₃);¹H−nmr (a small amount
CH₃CDCl including OD₃) is 1.3(s, 9H, t-bu
chill), 6.8(s, 1H) and 7.5(s, 1H)-imi
dazole proton, and 5.0(s,2H,CH ₂C₆
H_Five) included. Reference example 12 N-Alpha-((2R,4S,5S-6-shiku
lohexyl-5-[N-(t-butoxycarbonylate)
-Phenylalanyl)histidyl]amino-
4-hydroxy-2-(2-methylpropyl)
hexanoyl))-lysyl-phenylalanine di
acetate Product of Reference Example 11 (70 mg) and 10% Pd/C
(70mg) with 10ml of 4:1 methanol:acetic acid.
The mixture was hydrogenated at 4 atmospheres for 2 hours. filter catalyst
The filtrate was co-dispersed with toluene.
to give the title product as a white powder, 73 mg;
tlc Rf (1:1:1:1:ethyl acetate:butano
Le: Butanol: Acetic acid: H₂O), Rf 0.0 (1:19
Methanol: CHCl₃);¹H−nmr(DMSO−d₆)
contains 1.3(s,9H,t-butyl), benzene
Le CH₂No protons, aromatic at 6.8−7.8ppm
Contains protons; melting point, decomposes above 200°. Reference example 13 2R,4S,5S-6-cyclohexyl-5-
(t-butoxycarbonylamino)-4-hydro
xy-2-(2-methylpropyl)-N-methyl
Hexanamide (Method A) 2R,4S,5S-6-cyclohexyl-5-
(t-butoxycarbonylamino)-2-(2-methane)
tylpropyl)-gamma-hexanolactone (197
mg, 0.536 mmol, Reference Example 4) in 1 ml of water.
Dissolve and cool in an ice-water bath. CH cold solution₃
N.H.₃Irrigate with water for 3 minutes, stopper and leave at room temperature for 2 hours.
By then, tlc knows that the reaction is complete.
and showed. This mixture was solvent stripped and heated under high vacuum.
The title product forms a white fixed foam when dried under
204 mg; tlc Rf 0.31 (3:1 vinegar
ethyl acid: hexane). Reference example 14 2R,4S,5S-6-cyclohexyl-5-
Amino-4-hydroxy-2-(2-methylpropylene)
ropyru)-N-methylhexanamide hydrochloride Title product of Reference Example 13 (199 mg, 0.499 mmol)
) was dissolved in 4N HCl in dioxane (1 ml).
Se, N₂Stir for 25 minutes under the
TLC showed complete consumption of starting material. Next
Dry the solvent from the reaction mixture and dry under high vacuum.
If the title product is not completely dry,
Produced as a pale yellow powder, 185mg; tlc0.20
(18:2:1 CHCl₃: Ethanol : Acetic acid : TLC
Plate is NH before elution₃HCl exposed to steam
(changed the salt to the free base). Reference example 15 2R,4S,5S-6-cyclohexyl-5-
[N-(N-(t-butoxycarbonyl)pheni
lualanyl)-norleucyl]amino-4-hy
Droxy-2-(2-methylpropyl)-N-methane
Tylhexanamide The title product of Reference Example 14 (61.8 mg, 0.185 mmol)
), CH₂Cl₂(0.5ml), triethylamine (0.033
ml, 0.240 mmol), [N-(t-butoxycarbo
phenylalanyl]-norleucine (70 mg,
0.185 mmol), 1-hydroxybenzotriazo
(43 mg, 0.278 mmol) and dicyclohydrol (43 mg, 0.278 mmol)
Xylcarbodiimide (38mg, 0.185mmol)
were added sequentially to a 2 ml flask and the mixture was heated at 0° for 6 hrs.
Stir briefly and filter the mixture. filtrate, 2.5
ml CH₂Cl₂diluted with 2 x 3 ml 1N NaOH,
Wash with 1 x 3 ml brine and dry.
(MgSO_Four) When dissipated, an off-white powder and
Summer. This latter was chromatographed on 2g silica gel.
Tograph, CH₂Cl₂Medium 0.5%, 1%, 2
Gradient solution with 50 ml each of %, 4% and 6% ethanol.
away and contaminated with dicyclohexyl urea (DCU)
21.1 mg of white powder was obtained. This latter, first
The product and DCU isolated from the reaction mixture in
Chroma on 15 g of silica gel, including solids included.
Tograph, CH₂Cl₂1%, 2%, 3 inside
Gradient elution with 500 ml each of % and 4% ethanol
Ta. Combine and dissipate product fractions that do not contain DCU
The title product was obtained, 60.4 mg;¹H
−nmr(DMSO−d₆) 300mHz (ppm) is 0.9−
0.95(m, 9H, C(CH₃)₂and C-CH₃)；1.33
(s, 9H, boc); 2.56 (d, 3H, NCH₃, J = approx.
5Hz); 3.8, 4.18 and 4.32 (m, 1H, respectively);
4.62 (d, 1H, J = approx. 4Hz); 7.06, 7.46 and
7.95 (d, 1H, each, J = approximately 8 Hz), 7.68 (q,
1H, J = approximately 0.5Hz); 7.2-7.4 (m, aromatic). Reference example 16 2R,4S,5S-6-cyclohexyl-5-
(t-butoxycarbonylamino)-4-(2-
Tetrahydro-pyranyloxy)-2-(2-methane)
(tylpropyl)-N-methylhexanamide Title product of Reference Example 7 (211 mg, 0.449 mmol)
) was dissolved in 1 ml of dry THF and N₂below-
Cooled to 50°. While stirring, add N-methyl
Add morpholine (0.0545 ml, 0.494 mmol),
After 2-3 minutes, isobutyl chloroformate (0.0641
ml, 0.494 mmol) was added dropwise. Further at −50°
After 10 minutes, raise the temperature to -30° and add 0.068ml of THF.
Methylamine (37.6 mg, 1.21 mmol) in 20
It was added dropwise over a period of minutes. After 10 minutes, bring the mixture to ambient temperature.
Warm, dilute with 5 ml of ethyl acetate, and add 3 x 5 ml.
of saturated NaHCO₃, then 1 x 5 ml H₂Wash with O
and dried (MgSO_Four), diffused into oil,
245 mg. The latter was chromatographed on 5 g of silica gel.
Roughly eluted with 1:1 ether:hexane.
and tested by TLC to reveal the title product as a white solid foam.
Obtained as 102.2 mg; tlc Rf 0.2 and 0.25
(3:1 ether; hexane), which is a tetrahydride
1:1 epimer in lopyranyl ether side chain
is reflected. Reference example 17 2R,4S,5S-cyclohexyl-5-(t
-butoxycarbonylamino)-4-hydroxy
C-2-(2-methylpropyl)-N-methylh
Xanamide (Method B) The title product of Reference Example 16 (98.6 mg, 0.204 mmol)
2:1 acetic acid:H₂Dissolve in 1.5ml of O, 6
After stirring for an hour, dissipate and 2×CH₃C₆H_Five
was chased to obtain 99.2 mg of oil.
This latter was chromatographed on 5 g of silica gel.
200ml of 1:1, then 100ml of 3:1 acetic acid.
Eluted with ethyl:hexane and checked by TLC. Tooru
Combine and diffuse the bright product fractions to generate the title
Obtained as a white solid foam, 61.5 mg; tlc Rf
0.22 (2:1 ethyl acetate:hexane);¹H−nmr
(CDCl₃) 300mHz (ppm) is 0.9 (d, 6H),
Contains 1.46 (s, 9H), 2.82 (d, 3H); Reference example 13
was identical to the title product. This product (59.2 mg) was prepared by the method of Reference Example 14.
2 which is the same as the product of Reference Example 14R,4S,5S
-6-cyclohexyl-5-amino-4-hydro
xy-2-(2-methylpropyl)-N-methylhe
Changed to xanamide hydrochloride, 50.0 mg. Reference example 18 2R,4S,5S-6-cyclohexyl-5-
[N-alpha-(N-t-butoxycarbonyl
phenylalanyl)-N(imidazole)-(t-
butoxycarbonyl)histidyl]amino-4
-Hydroxy-2-(2-methylpropyl)-N
-Methylhexanamide By the method of Reference Example 15, CH₂Cl₂Ginseng in 1ml
Title product of Example 14 (47.3 mg, 0.141 mmol)
and N-alpha-[N-(t-butoxycarbo
phenylalanyl)-N(imidazole)-
(t-butoxycarbonyl)histidine (74.4 mg,
0.148 mmol) was converted to the title product. this
After filtering the reaction mixture, the filtrate was CH₂Cl₂dissipate
diluted with 1 ml of ethyl acetate and diluted with 2 x 1 ml of 1N
Wash with NaOH, 1x 1 ml brine and dry.
(MgSO_Four) and diffused to obtain 123.6 mg of foam.
Ta. Transfer this to a chromatograph on 4g of silica gel.
Rough, CH₂Cl₂Medium 0.5%, 1%, 2%, 4
Elute with 100 ml each of % and 6% ethanol,
Inspected by TLC. Combine the clear product fractions.
and dissipate to give the title product as a white solid foam.
tlc Rf 0.58 (18:2:1CHCl₃;
ethanol acetic acid). Reference example 19 2R,4S,5S-6-cyclohexyl-5-
[N-alpha-(N-t-butoxycarbonyl
phenylalanyl)-histidyl]amino-4
-Hydroxy-2-(2-methylpropyl)-N
-Methylhexanamide The title product from the previous reference example (81.1 mg, 0.104 mm
mol) in 1 ml of methanol, N₂under
So, about 5mg of K₂C.O.₃Stir for 1 hour with
Ta. K₂C.O.₃was neutralized with acetic acid and the mixture was allowed to dissipate.
dried, diluted with water, and ion-exchanged.
Desalt on resin (5 g, RP C-18) and 2x color.
2:3 methanol:H₂O, then 4x Ka
Elute with a ram volume of methanol. non-aqueous fraction
Combine, dissipate, dry under high vacuum, and
Obtained the title product as a white powder, 63.5 mg; tlc Rf
0.03(18:2:1CHCl₃;ethanol: acetic acid);¹H
−nmr(DMSO−d₆) 300mHz (ppm): 0.90
and 0.95 (double line, 6H in total, J = 7Hz, C
(CH₃)₂)；1.30(s,9H,C(CH₃)₃;2.56(d,
3H, J = approx. 5Hz, NCH₃); 3.72 (m, 1H); 4.14
(m, 1H); 4.47 (m, 1H); 7.53 (s, 1H, one
imidazolyl CH); 6.86, 7.12, 7.76 and
8.22 (m, each 1H, NH); 7.16-7.40 (m, aromatic
family). Reference example 20 2-[2R,4S,5S-6-cyclohexyl
-5-(t-butoxycarbonylamino)-4-
(2-tetrahydro-pyranyloxy)-2-
(2-methylpropyl)hexanoyl]-1,
2,3,4-tetrahydroisoquinoline The title product of Reference Example 7 (0.243 g, 0.517 mmol)
), tetrahydroisoquinoline (0.044ml,
0.069 g, 0.517 mmol) and 1-hydroxy
Benzotriazole (70 mg, 0.517 mmol)
5ml CH₂Cl₂and cooled to 0°. Jisik
lohexylcarbodiimide (107 mg, 0.517 mmol)
) and stir the mixture at room temperature for 18 hours.
Filter, strip the filtrate and dissolve the residue in 5 ml of acetic acid.
Triturated with ethyl and refiltered. the second filtrate, 1
x 5ml 5% HCl, 1 x 5ml saturated NaHCO₃,oh
and 1 x 5 ml brine and dry.
(MgSO_Four), dissipate the second residue, eluent
as CHCl₃Silica using 1% methanol in
Chromatographed on Kagel 36g, 229
mg;¹H−nmr (CDCl₃) Delta 1.4ppm (s, 9H,
C(CH₃)₃)including. Reference example 21 2-[2R,4S,5S-6-cyclohexyl
-5-amino-4-hydroxy-2-(2-methane)
tylpropyl)-hexanoyl]-1,2,3,
4-Tetrahydroisoquinoline hydrochloride By the method of Reference Example 14, the product of the previous Reference Example
(176 mg, 0.301 mmol) was completely dried.
153 mg;¹H-nmr (CD₃
OD) is delta 1.2ppm (6H, J=7, -CH
(CH₃)₂)including. Reference example 22 2-[2R,4S,5S-6-cyclohexyl
-5-(N-alpha-(N-t-butoxycal)
Bonyl-phenylalanyl)-N-(imidazo
)-(t-butoxycarbonyl)histidyl)
Amino-4-hydroxy-2-(2-methylpropylene)
(ropyl)-hexanoyl)-1,2,3,4-te
Trahydroisoquinoline N-methylmorpholine instead of triethylamine
Using the method of Reference Example 15, the previous Reference Example
of wet product (153 mg, assumed 0.301 mmol) and
and N-alpha-(N-t-butoxycarbonyl
phenylalanyl)-N(imidazole)-(t-butyl
carbonyl) histidine (186mg, 0.367mg)
limole) to change the title product. reaction mixture
Filter and filtrate with CHCl as eluent₃2.5% of
Chromatography on silica gel using methanol
to give the title product, 286 mg;¹H-
nmr (CDCl₃) is delta 1.4ppm (s, 9H, C
(CH₃)₃including. Reference example 23 2-[2R,4S,5S-6-cyclohexyl
-5-(N-(N-t-butoxycarbonylphene)
Nylalanyl)-histidyl)amino-4-hy
Droxy-2-(2-methylpropyl)-hexa
Noyl]-1,2,3,4-tetrahydroiso
quinoline The title product (286 mg) of the previous reference example was administered for 6 days.
4:1 acetic acid:H₂After stirring with 10ml of O,
Dissipate while toluene and remove the residue.
CHCl as eluent₃using 10% methanol in
Chromatographed on 20 g of silica gel,
Obtain the title product, the less polar of the products.
110mg;¹H−nmr (CDCl₃) is 1.4ppm (s,
9H,C(CH₃)₃including. Reference example 24 N-[2R,4S,5S-6-cyclohexyl
-5-(t-butoxycarbonyl phenylamine
-4-(2-tetrahydro-pyranyloxy)
c)-2-(2-methylpropyl)-hexanoyl
]Proline methyl ester Reaction time: 2 hours at 0° and 16 hours at room temperature
N- in place of triethylamine.
Using methylmorpholine and chromatography
CHCl as eluent in graphie₃1% methano in
Using the method of Reference Example 18, except using a
The product of Reference Example 7 (300 mg, 0.639 mmol)
) and proline methyl ester hydrochloride (106
mg; 0.639 mmol) to the title product, 215
mg;¹H−nmr (CDCl₃) is delta 1.4ppm (s,
9H,C(CH₃)₃); tlc Rf 0.7 (CHCl₃In
2.5%CH₃OH). Reference example 25 N-[2R,4S,5S-6-cyclohexyl
-5-(t-butoxycarbonyl phenylamine
-4-hydroxy-2-(2-methylpropyl)
)hexanoyl]proline methyl ester The product from the previous reference example (165 mg) was mixed with 4:1 vinegar.
Acid: H₂Stir in 10 ml of O for 18 hours and let it diffuse.
2 x 5 ml of toluene and CHCl₃1 x 5 ml
The title product that becomes moist as it aces is used as an oil.
obtained, 166 mg; tlc Rf 0.6 (CHCl₃Medium 2.5% CH₃
OH). Reference example 26 N-[2R,4S,5S-6-cyclohexyl
-5-amino-4-hydroxy-2-(2-methane)
thylpropyl)-hexanoyl]proline methoxy
luester The title product (166 mg, wet) from the previous reference example was
Combined with 3.7NHCl in oxane (1 ml). 2 o'clock
After a while, the mixture was evaporated and dissolved in 5 ml of ethyl acetate.
saturated NaHCO₃1 x 5ml then brine
Washed with 1 x 5 ml, dried (MgSO_Four)stop
and re-dissipated to give the title product,¹H−nmr
(CD₃OD) is delta 3.8ppm (s, 3H, CO₂CH
₃)including. Reference example 27 N-[2R,4S,5S-6-cyclohexyl
-5-(N-alpha-(N-t-butoxycal)
Bonyl-phenylalanyl)-N(imidazo
)-(t-butoxycarbonyl)histidyl)
Amino-4-hydroxy-2-(2-methylpropylene)
(propyl)-hexanoyl]proline methyl es
Tell Product of previous reference example (132 mg, 0.334 mmol),
N-alpha-(N-t-butoxy-carbonyl
phenylalanyl)-N(imidazole)-(t-butyl
carbonyl) histidine (168 mg, 0.334 mi)
rimole), 1-hydroxybenzotriazole
(45 ml, 0.334 mmol) and dicyclo-hexy
carbodiimide (69 mg, 0.334 mol) at 0°
5ml CH₂Cl₂Combine inside, CaCl₂by drying tube
After protection and stirring at this temperature for 18 hours,
Dissipate the medium and triturate the residue with 5 ml of ether.
Filtered. Residue obtained by sparging the ether filtrate
(230 mg) with CHCl as eluent₃1% CH in₃
Chromatography on silica gel using OH
to obtain the purified title product, 110 mg;¹H
−nmr(CDCl₃) is delta 1.4ppm (s, 9H, C
(CH₃)₃including. Reference example 28 2R,4S,5S-6-cyclohexyl-5-
[N-alpha-(N-t-butoxycarbonyl
-phenylalanyl)histidyl]amino-2
-(2-methylpropyl)-gamma-hexanola
Chthon Generate the title of the previous reference example using the method in reference example 25
Object, N-[2R,4S,5S-6-cyclohex
Sil-5-(N-alpha-(N-t-butoxyca)
Rubonylphenylalanyl)-N(imidazole)
-(t-butoxycarbonyl)histidyl)ami
No-4-hydroxy-2-(2-methylpropyl)
-hexanoyl] changed to proline methyl ester
110 mg;¹H-nmr is histidine t-butoxy
Indicated loss of carbonyl protecting group. This intermediate product
The composition is 1:24CH₃OH；CHCl₃silica using
When chromatographed on a gel, proline
formed the title product which lost the methyl ester,
51mg;¹H−nmr (CDCl₃) is delta 1.4ppm (s,
9H,C(CH₃)₃Contains methyl ester
does not include; ir (CDCl₃) is 1785cm^-1lactone
Contains a carbonyl group. Reference example 29 2R,4S,5S-6-cyclohexyl-5-
[N-alpha-(N-(t-butoxycarboni)
-Phenylalanyl)histidyl]amino-
4-hydroxy-2-(2-methylpropyl)
hexane amide Title product of previous reference example on 0.5 ml of methanol
(32.7 mg) of excess dry NH₃and perfused at 0°.
After about 10 minutes, allow the reaction mixture to evaporate and remove the residue.
Triturate with ether and filter the title product to white.
23 mg recovered as a colored solid;¹H−nmr (CDCl₃)
Delta 1.4ppm (s, 9H, C (CH₃)₃including. Reference example 30 N-[N-alpha-(t-butoxycarboni
)-N-epsilon-(benzyloxycarbo
lysyl]statin ethyl ester Does not require chromatographic purification
By the method of Reference Example 18, CH₂Cl₂25ml stati
Ethyl ester hydrochloride (0.467g, 0.00195mol)
), N-alpha-(t-butoxycarbonyl)
-N-epsilon-(benzyloxycarbonyl)
Lysyl (0.198g, 0.00195mol), N-methylmo
Luforin 0.215ml, 198mg, 0.00195mol), 1-H
Droxybenzotriazole (264mg, 0.00195mol)
) and dicyclohexylcarbodiimide
(403 mg, 0.00195 mol) as a solid foam of the title product.
Changed to droplet, 0.704g; tlc Rf 0.65 (9:1
CHCl₃:CH₃OH);¹H−nmr (CDCl₃) is delta
1.4ppm(s, 9H, C(CH₃)₃including. Reference example 31 N-[N-epsilon-(benzyloxycarbo
lysine]statin ethyl ester hydrochloride Product of previous reference example (0.805g, 0.00142mol)
for 2 hours in 10 ml of 3.7N HCl in dioxane.
Stir, dissipate and triturate the residue with ether.
Summed and filtered to obtain title generation address, 0.650g;
¹H-nmr (CD₃OD) is delta 5.2ppm (s, 2H,
Benzyl-CH₂)including. Reference example 32 N-[N-alpha-(2R,4S,5S-6-
Cyclohexyl-5-(t-butoxycarbonylate)
(ruamino)-4-(2-tetrahydro-pyranyl)
oxy)-2-(2-methylpropyl)hexano
yl)-N-epsilon-(benzyloxycal)
(bonyl)lysyl]-statin ethyl ester CHCl as eluent₃Using 0.5% methanol in
The product of the previous reference example by the method of Reference Example 27
(332 mg, 0.662 mmol) and production of Example 8
(311 mg, 0.662 mmol) was chromatographed.
300 mg of the title product;¹H−nmr
(CDCl₃) is delta 1.4(s,9H,C(CH₃)₃)of
include. Reference example 33 N-[N-alpha-(2R,4S,5S-6-
Cyclohexyl-5-(t-butoxycarbonylate)
Ruamino)-4-hydroxy-2-(2-methyl
propyl)hexanoyl)-N-epsilon-
(Benzyloxycarbonyl)lysyl]-Stati
ethyl ester By the method of Reference Example 25, the product of the previous Reference Example
(300 mg) to the title product and 2 x 10 ml of toluene.
dried under high vacuum, 200 mg;¹H
−nmr(CDCl₃) is delta 1.4ppm (s, 9H, C
(CH₃)₃)including. Reference example 34 N-[N-alpha-(2R,4S,5S-6-
cyclohexyl-5-amino-4-hydroxy
-2-(2-methylpropyl)hexanoyl)-
N-epsilon-(benzyloxycarbonyl)
Lysyl]-statin ethyl ester Product of previous reference example (200mg) and dioxa
Stir 10ml of 3.7N HCl in the bottle for 2 hours,
Dissipate, triturate the residue with ether and filter.
More white solid was collected. 1:19CH₃OH:
CHCl₃35g of these solids using
HCl-free due to chromagraphies on the tube
The title product was obtained, 52 mg;¹H-nmr (CD₃O.D.)
is delta 5.2(s, 2H, benzyl-CH ₂) including
nothing. Reference example 35 N-[[N-Alpha-[2R,4S,5S-6
-cyclohexyl-5-((N-alpha-(N
-t-butoxy-carbonyl phenylalani
)-N(imidazole)-(t-butoxycarboxylate)-N(imidazole)-(t-butoxycarbo
histidyl))amino-4-hydroxy
-2-(2-methylpropyl)hexanoyl]-
N-epsilon-[benzyloxycarbonyl]
Lysyl]] Statin ethyl ester 2 equivalents of 1-hydroxybenzotriazole
using and CHCl as eluent₃In
2.5% CH₃Using OH, except for the previous actual
Example product (52 mg, 0.0709 mmol) and N
-Alpha-(N-t-butoxycarbonylphene)
Nylalanyl)-N(imidazole)-(t-butoxy
(cycarbonyl)histidine (36mg, 0.0709mol)
to the chromatographed title product.
28mg;¹H−nmr (CDCl₃) is delta 1.4ppm
(s,9H,C(CH₃)₃) including; hplc retention time
10.28 min (reversed phase C18 column, 1:1 at 3 ml/min)
Acetonitrile: using PH2.1 phosphate buffer
elution). Reference example 36 N-[[N-Alpha-[2R,4S,5S-6
-cyclohexyl-5-(N-butoxycarbo
Nilphenylalanyl-histidyl)amino-
4-hydroxy-2-(2-methylpropyl)
hexanoyl]-N-epsilon-[benzilo
[oxycarbonyl]lysyl]]statin ethyl ethyl
Stell By the method of Reference Example 25, the product of the previous Reference Example
(28 mg) was converted into the title product, 22 mg;¹H-
nmr (CDCl₃) is imidazole t-butoxylic
demonstrated loss of the bonusyl protecting group or residual t-
Delta 1.4ppm (s,
9H) included. Reference example 37 N-[[N-Alpha-2R,4S,5S-6-
cyclohexyl-5-(N-t-butoxycal
Bonylphenylalanyl-histidyl)amino
-4-hydroxy-2-(2-methylpropyl)
hexanoyl [lysyl]]statin ethyl s
Tell The product (22 mg) of the previous reference example was added to 10% Pd/C22
hydrogenated for 3 hours at atmospheric pressure in 2 ml of ethanol on
did. Filter the catalyst over diatomaceous earth and add 2x ethanol.
It was collected by washing with 1 ml. combined filter
Dissipate the liquid and washing liquid to obtain the glass and minimize the amount
A white filter is produced by trituration with ether.
13 mg, converted into a liquid powder;¹H−nmr (CDCl₃)
is delta 1.5ppm (s, 9H, C (CH₃)₃)including. Reference example 38 2R,4S,5S-6-cyclohexyl-5-
(t-butoxycarbonylamino)-2-benzi
Le-gamma-hexanolactone N₂Below, in dry THF (7.4 ml, distilled from K)
Dry di(isopropyl)amine (2.48ml, 0.0177
Mol; CaH₂(distilled from ) was cooled to 0°. butyl
11.0 ml of lithium (1.62 M in hexane,
0.0177 mol) was added dropwise over 5 minutes. 15 minutes at 0°
After stirring, the mixture was cooled to −78° and dried.
4 of the smaller polarity of Reference Example 2 in 3.7ml of THF
The S, 5S -title product (2.30 g, 0.0074 mol) was added over 5 minutes. After stirring for an additional 30 minutes at -78°, benzyl bromide (0.923 ml, 0.0078 mol) in 3.7 ml dry THF was added dropwise over 5 minutes.
After 1 hour at −78°, the reaction was stopped by adding 10 ml of saturated NH ₄ Cl, warmed to room temperature, diluted with 25 ml of ether, and the layers were separated. organic layer,
Washed with 2 x 15ml of saturated _NaHCO3 , dried ( _MgSO4 ) and stripped to an oil (3.29g). This oil was chromatographed on 150 g of silica gel eluting with 2.5 liters of 1:9 ethyl acetate:hexane and examined by TLC. The clear product fractions were combined and stripped to give the purified title product as a white oily solid, 1.46 g; tlc Rf 0.60 (1:
1 ethyl acetate:hexane). The more polar starting material (0.91 g of 70% pure yellow oil, same system with tlc
Rf 0.4) was also recovered from this column. Reference example 39 2 R , 4 S , 5 S -6-cyclohexyl-5-
(t-Butoxycarbonylamino)-4-hydroxy-2-benzyl-N-methylhexanamide The title product (157 mg) of the previous reference example was dissolved in 2 ml of
Dissolved in CH ₃ OH, cooled to 0° and diluted with CH ₃ NH ₂
perfused for minutes. Stopper the flask and let it cool to room temperature.
Left for 1.5 hours, sparged with 2 x 2 ml of ether and dried under high vacuum to give the title product as a white solid foam, 164 mg; tlc
Rf 0.17 (1:1 ethyl acetate:hexane), Rf 0.29
(3:1 ethyl acetate:hexane); ¹ H-nmr
(CDCl ₃ , 300MHz is the expected C(CH ₃ ) ₃
(s.9H), N-CH ₃ (d, 3H), aromatic resonance and t-butoxycarbonyl-NH (d, 1H) peaks. Reference example 40 2 R , 4 S , 5 S -6-cyclohexyl-5-
Amino-4-hydroxy-2-benzyl-N-
Methylhexanamide hydrochloride The product of the previous reference example (159 mg) was dissolved in 2 ml of 4N HCl in dioxane, stirred under N ₂ for 0.5 h, allowed to evaporate, and chased with 3 x 2 ml of ether to remove the remaining residue. The material was dried under high vacuum to give the title product as a pale yellow powder, 135 mg;
tlc 0.20 (18:2:1 _CHCl3 :ethanol:acetic acid; spotted plates were exposed to _NH3 to neutralize HCl salts before elution). Reference example 41 2 R , 4 S , 5 S -6-cyclohexyl-5-
[N-alpha-(N-t-butoxycarbonylphenylalanyl)-N(imidazole)-(t-
butoxycarbonyl)histidyl]amino-4
-Hydroxy-2-benzyl-N-methylhexamide Title product of previous reference example (123 mg, 0.333 mmol), _CH2Cl3 (1 ml), triethyl-amine (0.060 ml, 0.433 mmol), N- _alpha- (N
-t-butoxycarbonylphenylalanyl)-
N(imidazole)-(t-butoxycarbonyl)
Histidine (176 mg, 0.350 mmol), 1-hydroxy-benzotriazole (84 mg, 0.550 mmol) and dicyclohexylcarbodiimide (72 mg, 0.350 mmol) were combined sequentially at 0° and stirred at this temperature for 16 h under _N2 . by-product
DCU was recovered by filtration and washing with 3 ml CH ₂ Cl ₂ . The combined filtrate and washing liquid are
After washing with 2 x 3 ml of 1N NaOH, brine 1 x 3
Wash with ml, dry ( _MgSO4 ) and diffuse
342 mg of a pale solid was chromatographed on 20 g of silica gel, 0.5%, 1% in CH ₂ Cl ₂
%, 2%, 4% and 6% ethanol each 250ml
Gradient elution with tlc Rf 0.55 (18:2:1
_CH2Cl3 : _ethanol :acetic acid). Reference example 42 2 R , 4 S , 5 S -6-cyclohexyl-5-
(t-butoxycarbonylphenylalanyl-
histidyl)-amino-4-hydroxy-2-
Benzyl-N-Methylhexanamide The title product (147 mg) of the previous reference example was stirred with 1.5 ml of 4: ₁ acetic acid:H 2 O under N ₂ for 7 hours, then stirred with 3 x 3 ml of ether. Trapping and drying under high vacuum gave the title product as a white powder. 135mg; tlc Rf0.05
(18:2:1 _CH3Cl3 : _ethanol :acetic acid); ^1H
−nmr (DMSO−d ₆ ), 250MHz is delta (ppm):
1.32 (s, 9H, t-butyl): 3.72, 4.15 and
4.48 (3m, 1H, respectively); 6.87 and 7.52 (s, 1H,
imidazolyl CH); 7.0-7.35 (m, aromatic), 7.40, 7.62 and 8.17 (3d, 1H, respectively). Reference example 43 2 R , 4 S , 5 S -6-cyclohexyl-5-
(t-Butoxycarbonylamino-4-hydroxy-2-(cyclohexylmethyl)-N-methylhexanamide) The title product (83.1 mg) of Reference Example 39 was dissolved in methanol.
10% Rh/C45mg in 10ml under 4 atm _H2
Hydrogenated for 1.5 hours. The catalyst was recovered by filtration over diatomaceous earth and washing with methanol. The combined filtrate and washings were stripped, chased with 2 x 5 ml ether and dried under high vacuum to give the title product as a white solid foam, 76.9 mg; ¹ H-nmr (CDCl ₃ ,
300MHz) contains a C(CH ₃ ) ₃ singlet, no phenyl protons, and an N-CH ₃ doublet;
tlc Rf 0.32 (3:1 ethyl acetate:hexane). Reference example 44 2 R , 4 S , 5 S -6-cyclohexyl-5-
Amino-4-hydroxy-2-(cyclohexyl-methyl)-N-methylhexanamide hydrochloride The title product (75 mg) of the previous reference example was converted to the title product according to the method of Reference Example 40, 64.1 mg ;tlc
Rf 0.8 (18:2:1 _CHCl3 :ethanol:acetic acid; spotted plates were neutralized with _NH3 vapor before elution). Reference example 45 2 R , 4 S , 5 S -6-cyclohexyl-5-
[N-alpha-(N-t-butoxycarbonylphenyl-alanyl)-N(imidazole)-(t
-butoxycarbonyl)histidyl]amino-
4-hydroxy-2-(cyclohexylmethyl)
-N-Methylhexanamide By the method of Reference Example 41, the product of the previous reference example (59.1 mg, 0.158 mmol) was converted to a white solid foam of the title product which was chromatographed, 76.4
mg, tlc Rf 0.60 (18:2:1 _CHCl3 :ethanol:acetic acid); ^1H -nmr (DMSO- _d6 , 250MHz)
Contains two C(CH ₃ ) ₃ singlets centered at 1.6 ppm. Reference example 46 2 R , 4 S , 5 S -6-cyclohexyl-5-
(t-butoxycarbonylphenylalanyl-
histidyl)amino-4-hydroxy-2-
(Cyclohexylmethyl)-N-methylhexanamide The product of the previous reference example (71.5 mg) was converted into the title product, which is a white powder, by the method of Reference Example 42.
59.1mg; tlc Rf 0.05 (18:2: _1CHCl3 :ethanol:acetic acid), ^1H -nmr (DMSO- _d6 , 250MHz) is:
1.32 ppm C( CH3 ) ₃ singlet, 2.58 ppm _N-
Contains a CH ₃ doublet and an imidazole C-H singlet at 6.86 and 7.52 ppm. Reference example 47 2 R , 4 S , 5 S -6-cyclohexyl-5-
(t-Butoxycarbonylamino)-4-hydroxy-2-(2-methylpropyl)-N-benzylhexanamide The title product of Reference Example 4 (150 mg, 0.408 mmol), dry toluene (2 ml), benzylamide (0.29 ml, 5 eq), acetic acid (0.023 ml, 1 eq),
Stir under N ₂ and gradually heat to 90° and hold at this temperature for 6 hours. The reaction mixture was cooled, diluted with 3 ml ethyl acetate, washed with 2 x 3 ml 1N HCl and 1 x 3 ml brine, dried (MgSO ₄ ) and stripped to give the title product as a white oily foam, 187 mg. ;tlc Rf 0.58 (2:1
ethyl acetate:hexane). Reference example 48 2 R , 4 S , 5 S -6-cyclohexyl-5-
Amino-4-hydroxy-2-(2-methylpropyl)-N-benzylhexanamide hydrochloride By the method of Reference Example 40, the product of the previous Reference Example (172 mg) was converted into the title product, which is a white powder. 146 mg; tlc Rf 0.30 (18:2:1 _CHCl3 :ethanol:acetic acid, the spotted plate was washed before elution.
(exposed to _NH3 vapor). Reference example 49 2 R , 4 S , 5 S -6-cyclohexyl-5-
[N-alpha-(N-t-butoxycarbonyl-phenylalanyl)-N(imidazole)-(t
-butoxycarbonyl)histidyl]amino-
4-hydroxy-2-(2-methylpropyl)-
N-Benzylhexanamide By the method of Reference Example 41, the product of the previous Reference Example (136 mg, 0.331 mmol) was converted to a white solid foam of the title product which was chromatographed, 167
mg; tlc Rf 0.60 (18:2:1 _CHCl3 :ethanol:acetic acid). Reference example 50 2 R , 4 S , 5 S -6-cyclohexyl-5-
(t-butoxycarbonylphenylalanyl-
histidyl)amino-4-hydroxy-2-
(2-Methylpropyl)-N-benzylhexanamide By the method of Reference Example 42, the product of the previous Reference Example was converted to the title product, which is a white, solid foam, 146 mg; tlc Rf 0.18 (18: 2: _1CHCl3 :ethanol:acetic acid); ^1H -nmr (DMSO- _d6 ), 250M
Hz) is delta (ppm); 0.83 and 0.90 (d, 3H,
respectively, J=6Hz, C(CH ₃ ) ₂ ; 1.32(s, 9H, C
(CH ₃ ) ₃ ); 3.72, 4.16 and 4.50 (m, 1H,
each); 4.28 (m, 2H, NCH ₂ ); 6.86 (br, 1H),
7.52 (s, 1H); 7.1-7.4 (m, 11-13H, aromatic and NH); 8.25 (d, 1H, NH); 8.36 (t,
1H) included. Reference example 51 2 R , 4 S , 5 S -6-cyclohexyl-5-
(t-Butoxycarbonylamino)-4-hydroxy-2-(2-methylpropyl)-N,N-dimethylhexanamide The title product of Reference Example 4 (595 mg) was dissolved in 5 ml of methanol and cooled to 10°. The tube was irrigated with dimethylamine for 3 minutes, stoppered, and left at room temperature for 1.5 hours. TLC showed 80% conversion. This solution was then perfused with dimethylamine for 2 minutes at room temperature, capped again and left for 18 hours, at which point
TLC showed complete conversion. Dissipate this solution and
The residue was dried under high vacuum to give the title product as a white solid foam, 112 mg; tlc Rf 0.31
(3:1 ethyl acetate:hexane); ms is 281.0,
Contains peaks at 230.9, 212.0, 186.1, 131.0, 100.0, 80.9, 68.9 and 57.0. Reference example 52 2 R , 4 S , 5 S -6-cyclohexyl-5-
Amino-4-hydroxy-2-(2-methylpropyl)-N,N-dimethylhexanamide hydrochloride The product of the previous reference example (107 mg) was prepared as an off-white solid foam by the method of Reference Example 40. The title product was modified, 94.7 mg; tlc Rf 0.16 (18:
2:1 _CHCl3 :ethanol:acetic acid; spotted plates were exposed to _NH3 vapor before elution); ms
are 313.3, 256.1, 215.2, 186.1, 156.0, 143.1,
Includes 126.1, 111.1, 100.1, 83.0 and 69.1. Reference example 53 2 R , 4 S , 5 S -6-cyclohexyl-5-
[N-alpha-(N-t-butoxycarbonylphenylalanine)-N(imidazole)-(t-
butoxycarbonyl)histidyl]amino-4
-Hydroxy-2-(2-methylpropyl)-
N,N-Benzylhexanamide According to the method of Reference Example 41, and in addition using 250 ml each of 8%, 12%, 20% and 50% ethanol in CH ₂ Cl ₂ for chromatographic elution, the previous reference The example product (87 mg, 0.249 mmol) was converted to the title product as a white solid foam, 107 mg; tlc
Rf 0.52 (18:2: _1CHCl3 , ethanol:acetic acid);
¹ H−nmr (DMSO−d ₆ , 250MHz) is a
( _CH3 ) ₃ single plug, including aromatic and imidazole peaks; ms 511.4, 357.3, 329.1,
301.2, 257.0, 211.1, 165.1, 136.0, 120.1,
110.0, 91.0, 57.1, 41.0. Reference example 54 2 R , 4 S , 5 S -6-cyclohexyl-5-
(t-butoxycarbonylphenylalanyl-
histidyl)amino-4-hydroxy-2-
(2-Methylpropyl)-N,N-benzylhexamide The product of the previous reference example was purified by the method of Reference Example 42 to give the crude title product, 78.2 mg, containing considerable melting point (mp) impurities by hplc. , changed to . This crude product was purified by preparative hplc on a 5 g reverse phase C-18 column.
1:1, 3:1, 17:3, and 100:
Eluted with 4 times the volume of each column of 0 methanol:H ₂ O. The impurity was eluted with this 3:1 eluent, while the purified title product was eluted with a 17:3 eluent and isolated by dispersion as a white powder, 46.0 mg; tlc Rf 0.02 (18: 2:1 _CHCl3 :ethanol:acetic acid); hplc retention time 5.87 minutes (C-8
4.6mm x 25cm column, 1.5ml/min 3: _2CH3CN :
elution with PH2.1 phosphate buffer); ms.322.4, 311.1,
283.1, 218.9, 165.1, 131.0, 110.1, 97.0, 91.0,
69.1, 59.1, 43.0; ¹ H−nmr (DMSO−d ₆ , 250M
Hz) is delta 1.32 (s, 9H, C(CH ₃ ) ₃ , 2.85 and 3.07 (2s, 6H, N(CH ₃ ) ₂ ), 6.92 and 7.52 (2s,
2H, imidazole C-H). Reference example 55 5-[2 R ,4 S ,5 S -6-cyclohexyl-5-(t-butoxycarbonylamino)-4-
(2-tetrahydro-pyranyloxy)-2-
(2-Methylpropyl)hexanoyl]amino-1-(benzyloxycarbonylamino)pentane The title product of Reference Example 7 was obtained by the method of Reference Example 30, except that ether was used instead of ethyl acetate in the isolation. (160 mg, 0.341 mmol) and the title product of Preparation 3 (80.5 mg, 0.341 mmol) were converted to the title product, 222 mg; ¹ H-
nmr (CDCl ₃ ) is delta 1.4 ppm (s, 9H, C
Contains ( _CH3 ) ₃ . Reference example 56 5-[ 2R , 4S , 5S -6-cyclohexyl-5-(t-butoxyamino)-4-hydroxy-2-(2-methylpropyl)hexanoyl]
Amino-1-(benzyloxycarbonylamino)pentane The product of the previous reference example (221 mg) was converted to the title product by the method of Reference Example 25, 193 mg; ¹ H-
nmr (CDCl ₃ ) delta 1.4ppm (s, 9H, C (CH ₃ ) ₃
including. Reference example 57 5-[ 2R , 4S , 5S -6-cyclohexyl-5-amino-4-hydroxy-2-(2-methylpropyl)hexanoyl]amino-1-
(Benzyloxycarbonylamino)pentane The product of the previous reference example (193 mg) was stirred with 10 ml of 3.7N HCl in dioxane for 2 hours,
Trapping and trituration of the residue with ether gave the title product, 144 mg; ¹ H-nmr (CD ₃ OD) containing delta 5.2 ppm (s, 2 H, benzyl CH ₂ ). Reference example 58 5-[[2 R ,4 S ,5 S -6-cyclohexyl-5-[N-alpha-N(t-butoxycarbonylphenyl-alanyl)-N(imidazole)
-(t-butoxycarbonyl)histidyl]amino-4-hydroxy-2-(2-methylpropyl)hexanoyl]]amino-1-(benzyloxycarbonylamino)pentane Using _CHCl3 as the eluent, Using the method, the product of the previous example was converted into the title product which was chromatographed, 25 mg; ¹ H-
nmr (CDCl ₃ ) is delta 1.4 ppm (s, 9H, C
Contains ( _CH3 ) ₃ . Reference example 59 5-[ 2R , 4S , 5S -6-cyclohexyl-5-(t-butoxycarbonylphenylalanyl-histidyl)-amino-4-hydroxy-2-(2-methylpropyl)hexanoyl]
Amino-1-(benzyloxycarbonylamino)pentane The title product (25 mg) of the previous reference example was converted into the title product by the method of Reference Example 42, and the residue after stripping was chased twice with toluene. , trituration of the residue with ether gave _the title product as a white solid, ¹¹ mg;
Contains 1.4ppm (s,9H,C( _CH3 ) ₃ . Using hydrogenation over Pb/C according to Reference Example 12,
This product was converted into 5-[2 R ,4 S ,5 S -6-cyclohexyl-5-(t-butoxycarbonylphenylalanyl-histidyl)-amino-4-hydroxy-2-(2-methylpropyl) Hexanoyl]Convert to aminopentylamine. Reference example 60 4-[2 R ,4 S ,5 S -6-cyclohexyl-5-(t-butoxycarbonylamino)-4-
(2-tetrahydro-pyranyloxy)-2-
(2-Methylpropyl)hexanoyl]benzyl aminobutyrate The title product of Reference Example 7 (160 mg, 0.341 mmol) and benzyl 4-aminobutyrate hydrochloride (66
mg, 0.341 mmol) were combined according to the method of Reference Example 22. At the end of this reaction period, the mixture is diffused, the residue is dissolved in 5 ml of ether, washed with 1 x 3 ml of saturated _NaHCO3 , 1 x 3 ml of 5% HCl and 1 x 3 ml of brine and re-sparged.
Obtained crude product, 172 mg. This is used as an eluent.
Chromatographed on 20 g silica gel using CHCl ₃ to give the purified title product, 107
mg; ¹ H−nmr (CDCl ₃ ) has a delta of 1.4 ppm (s,
Contains 9H, C(CH ₃ ) ₃ . Reference example 61 4-[2R,4S,5S-6-cyclohexyl-5-
(t-Butoxycarbonylamino)-4-hydroxy-2-(2-methylpropyl)hexanoyl]benzyl aminobutyrate The product (380 mg) of the previous reference example was converted to the title product by the method of Reference Example 42. 330 mg; ¹ H
-nmr includes delta 1.4ppm (s,9H,C( _CH3 ) ₃ . Reference example 62 4-[ 2R , 4S , 5S -6-cyclohexyl-5-amino-4-hydroxy-2-( Benzyl 2-methylpropyl)hexanoyl]aminobutyrate The product of the previous reference example (330 mg) in 10 ml of 3.7 N HCl in dioxane was allowed to stand for 2 hours. The mixture was allowed to evaporate and the residue was dissolved in 5 ml of ethyl acetate. saturated NaHCO ₃ and then brine, 5 each
Washed with 1 ml, dried (MgSO ₄ ) and redispersed to give the title product as an oil, 220 mg; ¹ H−
nmr (CD ₃ OD) is delta 5.2 ppm (s, 2H, CH ₂
C ₆ H ₅ ). Reference example 63 4-[[2 R ,4 S ,5 S -6-cyclohexyl-5-[N-alpha-(N-(t-butoxycarbonyl)phenyl-alanyl)-N(imidazole)-(t-butoxy Benzyl carbonyl)histidyl]amino-4-hydroxy-2-(2-methylpropyl)hexanoylamino]butyrate By the method of Reference Example 41, the product of the previous Reference Example (270 mg, 0.586 mmol) was combined to produce the title. Made things form. Following collection of DCU, the filtrate was stripped, dissolved in ether, refiltered, and the filtrate was stripped to 4.80 mg of crude product. The latter was chromatographed on 50 g silica gel using 2.5% methanol in _CHCl3 as eluent to give the purified title product, 132 mg; ¹ H
−nmr (CDCl ₃ ) is delta 1.4 ppm (s, 9H, C
Contains ( _CH3 ) ₃ . Reference example 64 4-[2 R ,4 S ,5 S -6-cyclohexyl-5-(N-(t-butoxycarbonyl)phenylalanyl-histidyl)amino-4-hydroxy-2-(2-methylpropyl)hexanoyl Amino]butyric acid The product (93 mg) of the previous reference example was added to 10% Pd/C23.
mg in 1:1 ethyl acetate; methanol 2.5 ml.
Hydrogenation was carried out at 4 atm for 2 hours. The catalyst was recovered by filtration and the filtrate was stripped to an oil. This was dissolved in ether, filtered and redispersed to give the title product, 62 mg; ¹ H-nmr ( _CD3OD )
contains delta 1.4ppm (s,9H,C( _CH3 ) ₃ . Reference example 65 2R , 4S , 5S- 6-cyclohexyl-5-
(t-butoxycarbonylamino)-2-[(2-
Naphthyl)methyl]-gamma-hexanolactone The title product of Reference Example 2 (0.295 g, 0.95 mmol) and 2-(bromomethyl)naphthalene (220 mg, 0.99 mmol) were converted to the crude title product by the method of Reference Example 38. (yellow oil 0.43g), and this
Chromatographed on 20 g of silica gel.
1:9 ethyl acetate; 700ml hexane, then 1:1
Eluted with ethyl acetate:hexane 300ml and checked by TLC. The clear title product was isolated from the central cut, 139 mg; Rf 0.62 (1:1 ethyl acetate:
hexane). Reference example 66 2 R , 4 S , 5 S -6-cyclohexyl-5-
(t-Butoxycarbonylamino)-4-hydroxy-2-[(2-naphthyl)methyl]-N-methylhexanamide The title product of the previous reference example (113 mg) in 2 ml of methanol was cooled to 0-5°. and convert this cold solution into _CH3
Purgated with _NH2 for 2 minutes, stoppered and left at ambient temperature for 2 hours, allowed to stand, chased with 2 x 2 ml of ether and dried under high vacuum to give the title product as a white foam. , 114mg; tlc Rf 0.15
(1:1 ethyl acetate:hexane), Rf 0.48 (18:
2:1 _CHCl3 :ethanol:acetic acid). Reference example 67 2 R , 4 S , 5 S -6-cyclohexyl-5-
Amino-4-hydroxy-2-[(2-naphthyl)methyl]-N-methylhexanamide hydrochloride 107.4 mg of the product of the previous Reference Example was purified by the method of Reference Example 14, which involves co-diffusion using ether. The title product turned into a solvent-moist yellow powder.
102.5 mg; Rf 0.2 (18:2:1 _CHCl3 :ethanol:acetic acid; loaded plates were previously neutralized with _NH3 vapor). Reference example 68 2 R , 4 S , 5 S -6-cyclohexyl-5-
(N-(N-alpha-(N-t-butoxycarbonyl)phenylalanyl)-N(imidazole)-(t-butoxycarbonyl)histidyl]
Amino-4-hydroxy-2-[(2-naphthyl)methyl]-N-methylhexanamide By the method of Reference Example 15, the title product of the previous Reference Example (96 mg, 0.229 mmol) and the title product of Preparation Example 1 were prepared. (121 mg, 0.240 mmol) was converted to an oily foam, 235 mg, of the crude title product. This was chromatographed on 5 g of silica gel and CH ₂
Elution was performed with 200 ml each of 0.5%, 1%, 2%, 4% and 6% ethanol in Cl ₂ and examined by tic. The clear product fraction was stripped to give the purified title product as a white foam, 124 mg; tlc Rf
0.6 (18:2:1 _CHCl3 :ethanol:acetic acid). Reference example 69 2 R , 4 S , 5 S -6-cyclohexyl-5-
(N-t-butoxycarbonylphenylalanyl-histidyl)-amino-4-hydroxy-
2-[(2-naphthyl)methyl]-N-methylhexanamide The title product of the previous reference example (116 mg) in 1.5 ml of methanol was stirred with K ₂ CO ₃ (1 mg) for 1 hour, then acetic acid Neutralized with 2 drops. the mixture,
Co-emission with ether gave the title product as a white powder, 101.4 mg; tlc Rf 0.08 (18:
2:1:ethanol:acetic acid); no msm ⁺ but mass peaks 311.1, 283.0, 257.0, 213.0, 192.1,
Includes 179.1, 141.0, 111.0, 97.1 and 57.1; ¹ H
−nmr (DMSO−d ₆ ) delta (300MHz), 1.30
(s, 9H, C(CH ₃ ) ₃ ), 2.46 (d, 3H, NCH ₃ ),
3.70, 4.14, 4.47 (3m, 1H each), 6.86, 7.62
(2s, 1H each), 7.10, 8.22 (2d, 1H each),
7.15−7.41 (m, aromatic), 7.48, 7.84 (2m, aromatic). Reference example 70 2 R , 4 S , 5 S -6-cyclohexyl-5-
[[N-[N-alpha-methyl-N-alpha-(N-t-butoxycarbonyl-phenylalanyl)-N(imidazole)-(t-butoxycarbonyl)]histidyl]]amino-4-hydroxy-2-( 2-(2-methylpropyl)-N,
N-Dimethylhexanamide By the methods of Reference Examples 15 and 68, the product of Reference Example 52 (0.534 g, 0.00153 mol) and N-alpha-methyl-N-alpha-(N-t-butoxycarbonylphenylar) were prepared. Nyl)-N(imidazole)-(t-butoxycarbonyl)]histidine (0.790 g, 0.00153 mol) was converted to an off-white foam of this chromatographed product, 0.394 g, tlc Rf 0.52 (18: 2:1:ethanol:acetic acid). Reference example 71 2 R , 4 S , 5 S -6-cyclohexyl-5-
[N-alpha-methyl-N-alpha-(N-
(t-butoxycarbonyl)-phenylalanyl)histidyl]amino-4-hydroxy-2
-(2-Methylpropyl)-N,N-dimethylhexanamide The product of the previous Reference Example (195 mg) was converted to the title product by the method of Reference Example 69 and preparative hpld on a RP C-18 column. and 4 column volumes of 1:
_1CH3OH : _H2O and 6 column volumes of 3: _1CH3
Purified by eluting with OH: _H2O . A clear product fraction was found in the latter eluent and was recovered as a white powder by dissipation, 153.6
mg; tlc Rf 0.09 (18:2:1 _CHCl3 :ethanol:acetic acid); ms m ⁺ is not present, but 575.3, 525.3,
511.3, 472.3, 442.2, 416.2, 399.2, 288.2,
371.2, 343.1, 291.0, 247.1, 186.1, 165.1,
Including 123.9, 109.0, 95.0, 69.1, 59.0, 43.0; ¹ H
−nmr (DMSO−d ₆ ) delta (300MHz) 0.78 (2d,
6H (CH ₃ ) ₂ rotamer), 3.0 (3s,
3H), 3.10, 3.76, 4.50, 4.95, 5.10, 6.92, 7.48,
7.60 (multiplet), 7.10−7.35 (broad multiplet, aromatic). Reference example 72 N-alpha-[ 2R , 4S , 5S- 6-cyclohexyl-5-(t-butoxycarbonyl)
Amino-4-(2-tetrahydropyranyloxy)-2-(2-methylpropyl)hexanoyl]-N-epsilon-[benzyloxy-carbonyl]lysine methyl ester Title product of Reference Example 7 (0.318 g, 0.00068 mol) ) was combined with N-epsilon-(benzyloxycarbonyl)lysine methyl ester hydrochloride (0.223 g, 0.00068 mol) according to the method of Reference Example 8. This crude product (0.261 g) was purified on silica gel using 99:1 _CHCl3 : _CH3OH as eluent.
Purified by chromatography on 30 g to give purified title product, 0.147 g; ¹ H-nmr
(CDCl ₃ ) contains delta 3.8 (s, 3H, OCH ₃ ). Reference example 73 H-alpha-[2 R , 4 S , 5 S -6-cyclohexyl-5-(t-butoxycarbonyl)
Amino-4-hydroxy-2-(2-methylpropyl)hexanoyl]-N-epsilon-[benzyloxycarbonyl]lysine methyl ester The title product of the previous reference example (0.131 g, 0.00018 mol) was dissolved in 4:1 acetic acid. : Stirring in 5 ml of H ₂ O for 16 hours, followed by sparging with toluene to give the title product as a white solid, 95 mg;
¹ H−nmr (CDCl ₃ ) is delta 3.8 (s, 3H,
Contains OCH ₃ ). Reference example 74 N-alpha-[ 2R , 4S , 5S -6-cyclohexyl-5-amino-4-hydroxy-2
-(2-methylpropyl)hexanoyl]-N-
Epsilon-[benzyloxycarbonyl]lysine methyl ester The title product (95 mg) of the previous reference example was dissolved at 0°C.
Dissolved in 2 ml of trifluoroacetic acid and kept for 1 hour, evaporated with toluene, partitioned the residue between 10 ml of ethyl acetate and 10 ml of saturated bicarbonate, and the aqueous layer was dissolved in 2 x 10 ml of fresh ethyl acetate. and the combined organic layers were washed with brine, dried over MgSO ₄ and stripped to give the title product as an oil, 80 mg; ¹ H-nmr.
(CDCl ₃ ) contains delta 3.8 (s, 3H, OCH ₃ ). Reference example 75 N-alpha-[[2 R ,4 S ,5 S -6-cyclohexyl-5-[N-alpha-(2-benzyl-3-phenylpropionyl)-N(imidazole)-(t-butoxy carbonyl)histidyl]amino-4-hydroxy-2-(2-methylpropyl)-hexanoyl]]-N-epsilon-[[benzyloxycarbonyl]]lysine methyl ester 99 as an eluent for chromatography:
1CHCl ₃ :Reference example except using CH ₃ OH
15 and 68, the product of the previous Reference Example (80 mg, 0.142 mmol) and the product of Preparation Example 9 (70 mg, 0.142 mmol) were combined to form the title product, 113 mg crude. , 80mg chromatographed; ¹ H-nmr (CDCl ₃ ) contains delta 1.6 (s, 9H, C(CH ₃ ) _3. Reference example 76 N-alpha-[[2 R , 4 S , 5 S -6-cyclohexyl-5-[N-alpha-(2-benzyl-3-phenyl-propionyl)histidyl]amino-4-hydroxy-2-(2-methylpropyl)hexanoyl]]-N-epsilon-[[benzyl [oxycarbonyl]]lysine methyl ester The product of the previous reference example (80 mg) was mixed with 4:1 acetic acid:
Dissolved in 3 ml of H ₂ O, kept for 24 hours, sparged with toluene, triturated with 3 ml of 1:1 ether:hexane and filtered to give the title product as a white powder, 40 ml; ¹ H−nmr
(CDCl ₃ ) shows no C(CH ₃ ) ₃ peak and a delta
Contains 3.8 (s, 3H, OCH ₃ ). Reference example 77 N-alpha-[[2 R , 4 S , 5 S -6-cyclohexyl-5-[N-alpha-(2-benzyl-3-phenylpropionyl)-histidyl]
Amino-4-hydroxy-2-(2-methylpropyl)hexanoyl]]lysine methyl ester Hydrogenation of the product of the previous reference example (40 mg) in 1 ml of methanol and 1 ml of acetic acid over 40 mg of 10% Pb/C for 1.5 hours. did. The catalyst was recovered by filtration and the filtrate was stripped to an oil and chased twice with toluene to give a white powder. Polish this with ether,
Filtration gave the title product, 18 mg; ¹ H-
nmr (CD ₃ OD) contains delta 3.8 ppm (s, 3H). This product is converted into the corresponding lysinamide derivative by the method of Reference Example 29. Reference Example 78 S-4-Methyl-2-(t-butoxycarbonylamino)pentanal N-(t-butoxycarbonyl)leucine methyl ester (28.0 g,
0.114 mol) to the title product, 21.7 g (88%) as a pale yellow oil, tlc Rf 0.36 (2:3 ethyl acetate; hexane); ¹ H-nmr ( _CDCl3 ) delta (90 MHz) 0.97 (d, J=6,6H), 1.1−1.8(m,
3H), 1.4 (s, 9H), 3.3-5.0 (m, 2H), 9.53
(s, 1H). Example 2 Ethyl 4RS , 5S -7-methyl-5-(t-butoxycarbonylamino)-4-hydroxy-2-octanoate using 3:17 and 1:4 ethyl acetate:hexane chromatography The product of the previous reference example (0.4 g, 0.048 mol) was converted to the title product by the method of Example 1, except using a gradient elution, 5.45 g, tlc Rf 0.40 (3:7 ethyl acetate:hexane); ir ( _CHCl3 ) 3438, 3340, 2233,
1711cm ^-1 ; ¹ H−nmr (CDCl ₃ ) delta (300MHz)
0.97 (t, J = 7, 6H), 1.34 (t, J = 6, 3H),
1.48 (s, 9H), 1.48 (m, 2H), 1.70 (m, 1H),
3.3-3.4 (m, 1H), 3.81-3.96 (m, 1H), 4.28
(q, J=7, 2H), 4.45−4.58 (m, 1H), 4.68
−4.78 (m, 1H). Analysis Calculated value as C ₁₆ H ₂₇ NO ₂ : C, 61.32; H, 8.68; N, 4.47 Actual value C, 61.38; H, 8, 58; N, 4.42 Reference example 79 4 S , 5 S -7-methyl -5-(t-Butoxycarbonylamino)-gamma-octanolactone Chromatography carried out according to the method of Reference Example 2, except for gradient elution with 2:3 to 1:0 ethyl acetate; hexane. The product of Example 2 was crystallized by changing the desired polar, relatively small ( 4S , 5S ) lactone product and trituration with hexane, 3.10 g (78%); mp.
76−77°; ir (CHCl ₃ ) 3439, 1775, 1711 cm ⁻¹ ; ¹ H
−nmr (CDCl ₃ ) delta (300MHz) 0.92 (d, J=
6, 6H), 1.44 (s, 9H), 1.28-1.81 (m, 3H),
2.06-2.32 (m, 2H), 2.48-2.58 (m, 2H), 3.79
-3.92 (wide width s, 1H), 4.42-4.58 (m, 2H). Analysis Calculated value as C ₁₄ H ₂₅ NO ₄ : C, 61.97; H, 9.29; N, 5.16 Actual value C, 62.15; H, 9.26; N, 5.12 Also relatively polar (4 R , 5 S ) lactone , isolated in much lower yield by chromatography and crystallized by hexane trituration, 0.68
g, melting point 113.5-116°C. Reference example 80 2 R , 4 S , 5 S -7-methyl-5-(t-butoxycarbonylamino)-2-(2-methyl-
2-propenyl)-gamma-octanolactone Method A The title product of the previous reference example (the less polar 4S , 5S -epimer;
0.51 g, 0.0019 mol) of the main title product, as well as its more polar 2 S , 4
S, 5S -epimer. Crude product (0.60
g) on a 4.5 x 20 cm silica gel column, 1:9, 3:17, 1:5,
Separation was achieved by elution with 1:4, 3:7 and 1:1 ether:hexane and collecting 23 ml fractions. From fractions 51-85, the purified title product was obtained, 0.21 g; mp 128-132°; [Alpha] ³⁰ _D -23.7° (C=0.529, _CH3OH )
The more polar 2S , 4S , 5S -epimer was isolated from fractions 89-120, 105 mg; ^1H -nmr
(CDCl ₃ ) contains delta 4.8 (2s, 2H, vinyl proton) and 1.75 (s, 3H, vinyl methyl); some of this epimer is crystallized by slow evaporation from CH ₂ Cl ₂ :hexane. When oxidized, needle-shaped crystals were obtained, melting point 99−101°. Method B Hexamethyldisilazane in 3.5 ml THF at 0°C
-78, prepared by dropwise addition of 5.1 ml (8.11 mmol) of a 1.6 M solution of n-butyllithium in hexane to 1.79 ml (1.39 g, 8.49 mmol)
A solution of 1.00 g (3.69 mmol) of the 4S , 5S -lactone of the previous reference example in 3 ml of THF was added dropwise to a suspension of lithium hexamethyldisilazide at .degree.
At the end of the addition, the mixture became clear and was stirred for an additional 15 minutes at -78°C. Next, THF2ml
0.548 g of freshly distilled methallyl bromide (4.06
A solution of 1 mmol) was added dropwise over 5 minutes, and the mixture was slowly warmed to -40°C over 2 hours.
The reaction was quenched with 2 ml of saturated NH ₄ Cl. After warming to room temperature, the reaction mixture was partitioned between 30 ml of ether and 30 ml of 10% citric acid. Separate the organic layer, add 10% citric acid (3 x 30ml) and sat.
Washed with NaHCO ₃ , dried (MgSO ₄ ) and evaporated to 1.11 g of a crude mixture of cis (2 S ) and trans (2 R ). The lactones were separated on 88 g of silica gel using ether-hexane (1:9 to 3:7) eluent. Trans ( 2S ) lactone with low polarity [tlc Rf 0.55 (1:
1 ether:hexane)] were combined and evaporated to give 0.613 g (51%) of a white solid, mp 132-135°C. Minor impurities (as indicated by TLC) were removed by trituration in hexane, yielding 0.562 g (47%) of analytically pure title 2.
R, 4S , 5S -lactone, melting point 133-135°C, was obtained. Crystals suitable for X-ray analysis were prepared by slow evaporation from hexane-methylene chloride. ¹ H−nmr (CDCl ₃ ) delta (250MHz) 0.90
(J, J=6,3H), 0.92 (d, J=6,3H),
1.42 (s, 9H), 1.70 (s, 3H), 1.92−2.15 (m,
2H), 2.26-2.39 (m, 1H), 2.57 (dd, J = 15 and 3, 1H), 2.72-2.88 (m, 1H), 3.77-3.90
(m, 1H), 4.34 (d, J=9, 1H), 4.33−4.51
(m, 1H), 4.70 (s, 1H), 4.81 (s, 1H); ¹³ C
−nmr (75mHz) delta 21.8, 23.0, 24.7, 28.3,
30.0, 37.9, 39.5, 41.8, 51.7, 79.8, 80.7,
112.8, 141.9, 156.0, 179.3, IR (CH ₂ Cl ₂ ) 3439,
1768, 1712, 1654 cm ⁻¹ ; [Alpha] _D −25.0° (c=0.5, CH ₃ OH). analysis
Calculated value as C ₁₈ H ₃₁ NO ₄ : C, 66.43; H,
9.60; N, 4.30. Actual value: C, 66.47; H, 9.59;
N, 4.27. Single crystal X-ray analysis demonstrated that the structural and stereochemical designations of these compounds were correct. The more polar cis (2S) lactone (tlc Rf
Fractions containing 0.44 1:11 ether; hexane) were combined and evaporated to give 39 mg (3%) of a white solid, mp 96-98 °C; ¹ H-nmr ( _CDCl3 ) delta (250 MHz) 0.92 (d, J-6, 6H), 1.43
(s, 9H), 1.72 (s, 3H), 2.02−2.14 (m, 1H),
2.23-2.36 (m, 1H), 2.60-2.87 (m, 2H), 3.74
-3.89 (m, 1H), 4.35-4.47 (m, 2H), 4.69 (s,
1H), 4.78 (s, 1H); ¹³ nmr (75MHz) delta
21.9, 22.0, 23.0, 24.8, 28.3, 30.7, 38.8, 38.9,
42.3, 50.1, 79.6, 80.4, 112.6, 142.0, 155.9,
178.7; IR (CHCl ₂ ) 3443, 1774, 1714, 1656cm ^-
1 ; [alpha] _D −0.6° (c=0.5, CH ₃ OH). analysis
Calculated value as C ₁₈ H ₃₁ NO ₄ : C, 66.43; H,
9.60; N, 4.30. Actual value: C, 66.94; H, 9.45;
N, 4.27. Reference example 81 2 R , 4 S , 5 S -7-methyl-5-(t-butoxycarbonylamino)-2-(2-methyl-
2-propyl)-gamma-octanolactone 10% A solution of 438 mg (1.35 mmol) of the title tictone from the previous reference example containing 44 mg of Pd/C in ethyl acetate (10 ml) was added to a Parr shaker.
Hydrogenation was performed on a Shaker apparatus at 50 psi for 2 hours. After filtration of the catalyst and evaporation of the weld, the title product
Obtained 437 mg (99%) as a white solid, mp 130-131°C. ¹ H−nmr (CDCl ₃ ) delta (300MHz)
0.84−0.97 (m, 12H), 1.41 (s, 9H), 1.86−
1.96 (m, 1H), 2.30-2.42 (m, 1H), 2.56-2.68
(m, 1H), 3.76−3.89 (m, 1H), 4.35 (d, J=
8, 1H), 4.45 (wide width t, 1H); ¹³ C−nmr (75M
Hz) delta 21.3, 21.8, 22.9, 23.0, 24.8, 26.1,
28.3, 31.0, 37.7, 40.5, 41.9, 51.7, 79.8, 80.5,
156.0, 180.3; IR (CHCl ₂ ) 3439, 1769, 1713cm ^-
1 ; [Alpha] _D −32.1° (C=1.0, CH ₃ OH). analysis
Calculated value as C ₁₈ H ₃₃ NO ₄ : C, 66.02; H,
10.16; N, 4.28. Actual value: C, 66.07; H,
10.03; N, 4.05. Various methods of the reference examples above convert this product into a homologous renin-inhibited product in which R ¹ is a 2-methylpropyl group rather than a cyclohexylmethyl group. Reference example 82 3-[2 R ,4 S ,5 S -6-cyclohexyl-5-(t-butoxycarbonylamino)-4-
(2-tetrahydropyranyloxy)-2-(2
-Methylpropyl)hexanoylamino]benzyl propionate The product of Reference Example 7 (0.30 g, 0.64 mmol) and beta-alanine benzyl ester hydrochloride (0.15 g, 0.70 mmol) were mixed at 0°C with 5 ml of CH ₂ Cl ₂
I matched it inside. With stirring, N-methylmorpholine (0.154 ml, 1.41 mmol) was added followed by diethyl cyanophosphonate (0.109 ml, 0.70 mmol). 16 hours at 0° and 1 at room temperature
After stirring for an hour, the mixture was allowed to dry and the residue was dissolved in ethyl acetate (20 ml), extracted with 2 x 20 ml of saturated _NaHCO3 , 1 x 20 ml of _H2O and 1 x 20 ml of brine, dried ( MgSO ₄ ), stripped to give 0.394 g of crude product, which was purified with 99:1 CHCl ₃ :CH ₃ as eluent.
Chromatographed on 45 g of silica gel using OH to yield the purified title product;
0.30g:tlc Rf 0.8 (19:1: _CHCl3 : _CH3OH ). Reference example 83 3-[2 R ,4 S ,5 S -6-cyclohexyl-5-(t-butoxycarbonylamino)-4-
Benzyl hydroxy-2-(2-methylpropyl)hexanoylamino]propionate The product of the previous reference example (300 mg) was mixed with 4:1 acetic acid:
Stirring with 10 ml of H ₂ O for 16 hours, stripping under high vacuum and sieving with toluene gave the title product, 273 mg. Reference example 84 3-[ 2R , 4S , 5S -6-cyclohexyl-5-amino-4-hydroxy-2-(2-methylpropyl)hexanoylamino)propionate benzyl hydrochloride Production of the previous reference example (273 mg) was dissolved in 3 ml of ether and 3.7N in dioxane at 0°C.
Treated with 3 ml of HCl. After 4 hours at 0°, the reaction was evaporated and triturated with ether-hexane to give the title product as a white solid, 248 mg. Reference example 85 3-[2 R , 4 S , 5 S -6-cyclohexyl-5-[N-alpha-(N-(t-butoxycarbonyl)-phenylalanyl)-N(imidazole)-(t-butoxycarbonyl) benzyl histidyl]amino-4-hydroxy-2-(2-methylpropyl)hexanoylamino)propionate 39:1 as eluent in chromatography
CHCl ₃ :Reference example except using CH ₃ OH
In the presence of N-methylmorpholine (0.155 ml, 1.413 mmol) and diethyl cyanophosphonate (0.11 ml, 0.707 mmol) using the method of Example 82, the product of the previous reference example (248 mg, 0.589 mmol) and Preparation Example 1 product (357 mg, 0.707
The purified title product was prepared by combining 0.225 g; tlc 0.5 (9:1
_CHCl3 : _CH3OH ). Reference example 86 3-[ 2R , 4S , 5S -6-cyclohexyl-5-[N-(t-butoxycarbonyl)phenylalanyl-histidyl]amino-4-hydroxy-2-(2-methylpropyl)hexanoyl Benzyl amino]propionate The product of the previous reference example (342 mg) was converted to the title product by the method of Reference Example 83, 305 mg; tlc
Rf 0.2 (9:1 _CHCl3 : _CH3OH ). Reference example 87 3-[ 2R , 4S , 5S -6-cyclohexyl-5-[N-(t-butoxycarbonyl)phenylalanyl-histidyl]amino-4-hydroxy-2-(2-methylpropyl)hexanoyl Aminopropionic acid The product of the previous Reference Example (0.305 g) was hydrogenated according to Reference Example 12 using 10 ml of pure methanol as solvent. Recovery of the catalyst followed by stripping of the filtrate gave the title product, 0.234 g; ¹ H-nmr
(CDCl ₃ ) contains delta 1.5 ppm (s, 9H). Reference example 88 2 R , 4 S , 5 S -6-cyclohexyl-5-
(t-butoxycarbonylamino)-4-hydroxy-2-(2-methyl-2-propenyl)-N
-Methylhexanamide trans( 2R ) title product of Reference Example 3 (0.20
g, 0.55 mmol) in 10 ml of CH ₃ OH,
Cooled to 0° C. and the solution was saturated with CH ₃ NH ₂ .
After 60 hours at 0°, the mixture was stripped and the solid residue was triturated with hexane to give the title product, 0.220 g. Reference example 89 2 R , 4 S , 5 S -6-cyclohexyl-5-
Amino-4-hydroxy-2-(2-methyl-
2-propenyl)-N-methylhexanamide The product of the previous reference example (0.220 g) was heated at -11°C.
The mixture was stirred with 2 ml of trifluoroacetic acid for 0.5 hour. The reaction mixture was stripped down and the residue was dissolved in 5 ml of ethyl acetate, washed with 5 ml of saturated NaHCO ₃ and 5 ml of brine, dried (MgSO ₄ ) and stripped to give the title product.
0.169g. Reference example 90 2 R , 4 S , 5 S -6-cyclohexyl-5-
[N-alpha-(N-t-butoxycarbonylphenylalanyl)-N(imidazole)-(t-
butoxycarbonyl)histidyl]amino-4
-Hydroxy-2-(2-methyl-2-propenyl)-propenyl N-methylhexanamide Product of previous reference example (169 mg, 0.57 mmol)
and the product of Preparation Example 1 (288 mg, 0.57 mmol)
were combined at 0° in 15 ml of CH ₂ Cl ₂ with the addition of 1-hydroxybenzotriazole (77 mg, 0.57 mmol) and dicyclohexylcarbodiimide (118 mg, 0.57 mmol). After stirring at ambient temperature for 18 hours, dicyclohexylurea was recovered by filtration, the filtrate was stripped and the resulting residue was dissolved in 10 ml of ethyl acetate and saturated.
Washed with 1 x 5 ml _NaHCO3 , 1 x 5 ml 5% HCl and 1 x 5 ml brine, dried ( _MgSO4 ), diffused (0.329 g), 40:1 _CHCl3 :
Chromatography on 35 g of silica gel using CH ₃ OH as eluent gave the purified title product, 0.129 g. Reference example 91 2 R , 4 S , 5 S -6-cyclohexyl-5-
[N-(t-butoxycarbonyl)phenylalanylhistidyl]amino-4-hydroxy-2
-(2-Methyl-2-propenyl)-N-methylhexanamide Reference example of finally triturating the solid product with ether
By the method of 83, the product of the previous reference example (0.129
g) into the title product, 75 mg; ¹ H-
nmr (CDCl ₃ ) contains delta 1.5 ppm (s, 9H). Reference example 92 4-[ 2R , 4S , 5S -cyclohexyl-5
-(t-butoxycarbonylamino)-4-(2
-tetrahydropyranyloxy)-2-(2-methylpropenylhexanoylamino]ethyl butyrate Product of Reference Example 6 (300 mg, 0.64 mmol) using the method of Reference Example 82 without chromatography
was combined with ethyl 4-aminobutyrate hydrochloride (118 mg, 0.70 mmol) to give the title product, 385
mg. Reference example 93 4-[2 R ,4 S ,5 S -6-cyclohexyl-5-(t-butoxycarbonylamino)-4-
Ethyl hydroxy-2-(2-methylpropyl)hexanoylamino]butyrate The product (0.385 g) of the previous reference example was added to Reference Example 83.
Treated with 4:1 acetic acid:H ₂ O according to
The lactone-containing product (330 mg) was isolated by lyophilization and the title product was eluted with 1:1 ether:hexane for the lactone and 19:1 for the title product.
161 mg isolated by chromatography on silica gel eluting with CHCl ₃ CH ₃ OH; tlc
Rf 0.6 (9:1 _CHCl3 : _CH3OH ). Reference Example 94 4-[ 2R , 4S , 5S -6-cyclohexyl-5-amino-4-hydroxy-2-(2-methylpropyl)-hexanoylamino]ethyl butyrate hydrochloride Production of the previous reference example (0.161 g) was dissolved in 5 ml of ether, cooled to 0°, 7.4N HCl in dioxane (3 ml) was added and the mixture was stirred at 0°C for 3.5
Stirring for an hour, then stripping and drying under high vacuum gave the title product, 0.141g, tlc Rf
0.1 (9:1 _CHCl3 : _CH3OH ). Reference example 95 4-[2 R ,4 S ,5 S -6-cyclohexyl-5-[N-alpha-(N-t-butoxycarbonyl)phenylalanyl-N(imidazole)
-(t-Butoxycarbonyl)histidyl]amino-4-hydroxy-2-(2-methylpropyl)hexanoylamino]ethyl butyrate The product of the previous reference example (140 mg, 0.323 mmol) and The product of Preparation 1 (179 mg, 0.355 mmol) was combined, isolated, and purified to give the title product, 136 mg. By the method of Reference Example 86, this product was converted into 4-[2 R ,4
S, 5S -6-cyclohexyl-5-[N-(t-
butoxycarbonyl)-phenylalanyl-histidyl]amino-4-hydroxy-2-(2-methylpropyl)hexanoylamino]-ethyl butyrate. Reference example 96 2 R , 4 S , 5 S -6-cyclohexyl-5-
(t-butoxycarbonylamino)-2-(p-
Chlorbenzyl)-gamma-hexanolactone The less polar 4S , 5S -product of Reference Example 2 (2.5 g, 0.008 mol) was prepared by the method of Reference Example 38.
and 4-chlorobenzyl bromide (1.81 g, 0.0088
tlc Rf 0.6 (3:1
hexane:ethyl acetate) 0.7 (2:1 hexane:
ethyl acetate containing 1% acetic acid). Reference example 97 2 R , 4 S , 5 S -6-cyclohexyl-5-
(t-butoxycarbonylamino)-2-(p-
methylbenzyl)-gamma-hexanolactone The 4 R ,5 S -product of Reference Example 2 ( 1.5g,
tlc Rf 0.55 (2:1
hexane:ethyl acetate), 0.75 (1:1 hexane:
Ethyl acetate). Reference example 98 2 R , 4 S , 5 S -cyclohexyl-5-(t
-butoxycarbonylamino)-2-(p-methoxybenzyl)-gamma-hexanolactone 4S , 5S of Reference Example 2 by the method of Reference Example 97
- product (3.79 g, 0.0118 mol) and p-bromide
Methoxybenzyl (2.61 g, 0.130 mol) was chromatographed to give the title product, a white gum, 1.06 g; tlc Rf 0.4 (3:1 hexane:ethyl acetate). Reference example 99 2 R , 4 S , 5 S -6-cyclohexyl-5-
(t-butoxycarbonylamino)-2-(3,
4-dichlorobenzyl)-gamma-hexanolactone The 4S , 5S -product of Reference Example 2 (2.0 g ，
0.0064 mol) and 3,4-dichlorobenzyl bromide (1.68 g, 0.007 mol) to the title product, a clear gum, 1.36 g; tlc Rf 0.22 (3:1 hexane:ethyl acetate), 0.9 (1:2 hexane:1
% acetic acid (ethyl acetate). Reference example 100 2 R , 4 S , 5 S -6-cyclohexyl-5-
(t-butoxycarbonylamino)-2-(o-
Chlorbenzyl)-gamma-hexanolactone The 4S , 5S -product of Reference Example 2 (2.0 g,
0.0064 mol) and o -chlorobenzyl chloride (1.44 g, 0.007 mol) to the title product, a clear gum, 1.51 g; tlc Rf 0.75 (3:1 hexane:ethyl acetate), 0.65 (2:1 Hexane: ethyl acetate containing 1% acetic acid). Reference example 101 2 R , 4 S , 5 S -6-cyclohexyl-5-
(t-butoxycarbonylamino)-2-(m-
Chlorbenzyl)-gamma-hexanolactone 4S , 5 of Reference Example 2 by the method of the previous Reference Example
S-product (2.0 g, 0.0064 mol) and m chloride
-Chlorbenzyl (0.94ml, 1.44g, 0.007mol)
was converted to a colorless gum of the title product which was chromatographed, 2.11 g; tlc Rf 0.4 (3:1 hexane: ethyl acetate), 0.75 (2:1 hexane: ethyl acetate with 1% acetic acid). Reference example 102 2 R , 4 S , 5 S -6-cyclohexyl-5-
(t-Butoxycarbonylamino)-4-hydroxy-2-(p-chlorobenzyl)-N-methylhexanamide The method of Reference Example 88 using a reaction time of 2 hours at ambient temperature and without hexane trituration. using,
The product of Reference Example 96 (1.0 g) was converted into the title product, white solid, 1.07 g; tlc Rf 0.75 (2:1
Hexane: ethyl acetate containing 1% acetic acid), 0.6
(18:2: _{1 CH2Cl2} : _C2H5OH :acetic _acid ). Reference example 103 2 R , 4 S , 5 S -6-cyclohexyl-5-
(t-Butoxycarbonylamino)-4-hydroxy-2-(p-methylbenzyl)-N-methylhexanamide 1:1 as eluent using the method of the previous reference example
Chromatographing the product on silica gel with hexane:ethyl acetate converted the product of Reference Example 97 (0.885 g) to the title product, 0.518
g; tlc Rf 0.1 (1:1 ethyl acetate:hexane),
0.55 (18:2:1 _CHCl3 : _C2H5OH :acetic _acid ). Reference example 104 2 R , 4 S , 5 S -6-cyclohexyl-5-
(t-Butoxycarbonylamino)-4-hydroxy-2-(p-methoxybenzyl)-N-methylhexanamide The product of Reference Example 98 (1.06 g) was prepared as an off-white solid by the method of Reference Example 3. tlc Rf 0.2 (2:1 ethyl acetate: 1% acetic acid in hexanes), 0.8 (9:
_1CH2Cl2 : _C2H5OH containing 1% concentrated _{NH4OH ) .} Reference example 105 2 R , 4 S , 5 S -6-cyclohexyl-4-
(t-butoxycarbonylamino)-4-hydroxy-2-(3,4-dichlorobenzyl)-N-
Methylhexanamide The product of Reference Example 99 (1.36 g) was changed into the present product, which is a white rubber, by the method of Reference Example 102.
1.39g; tlc Rf 0.25 (1:1 hexane:ethyl acetate containing 1% acetic acid), 0.55 (9: _{1 CH2Cl2} :1% conc.
_C2H5OH including _{NH4OH )} . Reference example 106 2 R , 4 S , 5 S -6-cyclohexyl-5-
(t-Butoxycarbonylamino)-4-hydroxy-2-(o-chlorobenzyl)-N-methylhexanamide Reference Example 102 was prepared using the method of Reference Example 102, except that a reaction time of 16 hours at ambient temperature was used. The product of Example 100 (1.51 g) was changed to the white rubber of this product,
1.56g; tlc Rf 0.09 (2:1 hexane: acetic acid containing 1% acetic acid. Reference example 107 2 R , 4 S , 5 S -6-cyclohexyl-5-
(t-Butoxycarbonylamino)-4-hydroxy-2-(m-chlorobenzyl)-N-methylhexanamide The product of Reference Example 101 (2.11 g) was converted into the title product by the method of Reference Example 102. Changed to white rubber,
1.68 g; tlc Rf 0.25 (2:1 hexane: ethyl acetate with 1% acetic acid), 0.5 (9: _{1 CH2Cl2} : _C2H5OH with 1% _acetic acid). Reference example 108 2 R , 4 S , 5 S -6-cyclohexyl-5-
Amino-4-hydroxy-2-(p-chlorobenzyl)-N-methylhexanamide hydrochloride The product of Reference Example 102 (1.07 g) was dissolved in 10 ml of 3.78 N HCl in dioxane for 10 min under _N2 . Stir and dissipate. Dissolve the residue with 10 ml of ether and 3
Combined and redispersed to yield the title product as a solid, 1.11 g; elc Rf 0.18 (18:2:1 CH _2
Cl2 : _C2H5OH :acetic _acid ). Reference example 109 2 R , 4 S , 5 S -6-cyclohexyl-5-
Amino-4-hydroxy-2-(p,methylbenzyl)-N-methylhexanamide hydrochloride The product of Reference Example 103 (0.454 g) was converted into the title product by the method of the previous reference example. 0.4g;
tlc Rf 0.1 (18:2:1 _CHCl3 : _C2H5OH :acetic _acid ). Reference example 110 2 R , 4 S , 5 S -6-cyclohexyl-5-
Amino-4-hydroxy-2-(p-methoxybenzyl)-N-methylhexanamide hydrochloride The product of Reference Example 104 (1.07 g) was converted to the title product in quantitative yield by the method of Reference Example 108. (weight yield 1.14g, 0.22g more than theory);
tlc Rf 0.2 (9:1 CH ₂ Cl ₂ : Contains 1% concentrated NH ₄ OH
_C2H5OH ) _. Reference example 111 2 R , 4 S , 5 S -6-cyclohexyl-5-
Amino-4-hydroxy-2-[3,4-dichlorobenzyl)-N-methylhexanamide hydrochloride The product of Reference Example 105 (1.38 g) was obtained in quantitative yield by the method of Reference Example 108. Changed to title product (weight 1.52g, 0.30g more than theory); tlc Rf
0.25 ( _CH2Cl2 : _C2H5OH9 :1 with 1% _{concentrated NH4OH} ) _. Reference example 112 2 R , 4 S , 5 S -6-cyclohexyl-5-
Amino-4-hydroxy-2-[o-chlorobenzyl)-N-methylhexanamide hydrochloride The product of Reference Example 106 (1.56 g) was converted to the title product by the method of Reference Example 108, 1.4 g;tlc
Rf 0.1 (9:1 CH ₂ Cl ₂ :C ₂ H ₅ with 1% acetic acid
OH). Reference example 113 2 R , 4 S , 5 S -6-cyclohexyl-5-
Amino-4-hydroxy-2-(m-chlorobenzyl)-N-methylhexanamide hydrochloride The product of Reference Example 107 (1.67 g) was converted to the title product in quantitative yield by the method of Reference Example 108. (0.23g more than theory); tlc 0.1 (9:1CH _2
Cl2 : _C2H5 containing 1% _acetic acid). Reference example 114 2 R , 4 S , 5 S -6-cyclohexyl-5-
[N-(t-butoxycarbonyl)phenylalanyl-N(imidazole)-(t-butoxycarbonyl)histidyl]amino-4-hydroxy-2-(p-chlorobenzyl)-N-methylhexanamide Reference Example 108 under nitrogen product (1.11g, 0.0023
mol) was stirred with 8 ml of CH ₂ Cl ₂ .
Sequentially triethylamine (0.42 ml, 0.0030 mol),
Product of Preparation Example 1 (1.20 g, 0.0024 mol), 1-
Hydroxybenzotriazole (0.583g,
0.0038 mol) and dicyclohexylcarbodiimide (0.495 g, 0.0024 mol) were added and the mixture was stirred for 24 hours. The reaction mixture was purified by CH ₂ Cl ₂
Dilute with 25 ml of 1N NaOH 2 x 12 ml and saturate
Wash with 1 x 12 ml of NaCl and dry (Na ₂ SO ₄ ).
Dissipated into foam, 1.31g. This latter was chromatographed on silica gel using 19:1 CH ₂ Cl ₂ :C ₂ H ₅ OH as eluent to give the purified title product, 0.70 g; tlc Rf 0.65
(18:2:1 _CH2Cl2 : _{C2H5OH :} acetic _acid ). Reference example 115 2 R , 4 S , 5 S -6-cyclohexyl-5-
[N-(t-Butoxycarbonyl)phenylalanyl-N(imidazole)-(t-butoxycarbonyl)histidyl]amino-4-hydroxy-2-(p-methylbenzyl)-N-methylhexanamide Combine the reagents at 0°C. Reference Example 109 was prepared using the method of the previous Reference Example, except for warming the reaction mixture after heating and using 5:1 CH ₂ Cl ₂ :C ₂ H ₅ OH as the eluent for the chromatography.
(0.4g) was converted into the title product,
0.60g; tlc Rf 0.9 (18:2:1 CH ₂ Cl ₂ :C ₂ H ₅
OH: acetic acid). Reference example 116 2 R , 4 S , 5 S -6-cyclohexyl-5-
[N-(t-butoxycarbonyl)phenylalanyl-N(imidazole)-(t-butoxycarbonyl)histidyl]amino-4-hydroxy-2-(p-methoxybenzyl)-N-methylhexanamide Chromatography eluent as 20:1
The product of Reference Example 110 (0.92 g, corrected for purity) was converted to the title product as a white solid by the method of the previous Reference Example using CH ₂ Cl ₂ :C ₂ H ₅ OH, 0.73 g; tlc Rf 0.7 (9:1CH ₂ Cl ₂ :C ₂ H ₅
OH), 0.85 (9:1 _CH2Cl2 : ₁ % concentrated _NH4OH )
_CH30H ). Reference example 117 2 R , 4 S , 5 S -6-cyclohexyl-5-
[N-(t-butoxycarbonyl)phenylalanyl-N(imidazole)-(t-butoxycarbonyl)histidyl]amino-4-hydroxy-2-(3,4-dichlorobenzyl)-N-methylhexanamide chromatography 24:1 as an eluent
The product of Reference Example 111 (1.22 g, corrected for purity) was converted to the title product by the method of Reference Example 115 using CH ₂ Cl ₂ :C ₂ H ₅ OH, 1.08 g; tlc Rf
0.75 (9:1 _CHCl3 : _C2H5OH ) _. Reference example 118 2 R , 4 S , 5 S -6-cyclohexyl-5-
[N-(t-butoxycarbonyl)phenylalanyl-N(imidazole)-(t-butoxycarbonyl)histidyl]amino-4-hydroxy-2-(o-chlorobenzyl)-N-methylhexanamide By the method of Reference Example 117 , the product of Reference Example 112 (1.33 g, corrected for purity) was chromatographed into a white solid of the title product,
1.02g; tlc Rf 0.55 (9:1 _{CH2Cl2 :} ethyl acetate containing 1% acetic acid). Reference example 119 2 R , 4 S , 5 S -6-cyclohexyl-5-
[N-(t-butoxycarbonyl)phenylalanyl-N(imidazole)-(t-butoxycarbonyl)histidyl]amino-4-hydroxy-2-(m-chlorobenzyl)-N-methylhexanamide By the method of Reference Example 117 , 1.68 g of the product of Reference Example 113 (1.45 g, corrected for purity) was chromatographed into the white gum of the title product;
tlc Rf 0.62 (9: _1CH2Cl2 : _C2H5 with 1% _{acetic acid}
OH). Reference example 120 2 R , 4 S , 5 S -6-cyclohexyl-5-
(t-butoxycarbonylphenylalanyl-
histidyl)-amino-4-hydroxy-2-
(p-chlorobenzyl)-N-methylhexanamide The product (0.25 g) of Reference Example 114 was added to 2 ml of glacial acetic acid.
and 0.5 ml of H ₂ O, stirred under N ₂ for 8 h, then allowed to dry, and the residue was triturated with 3 x 0.5 ml of toluene and then 3 x 5 ml of ether, re-sparging each time. Make slurry with 83
mg of crude product was obtained. This latter was prepared using 1:1 CH ₃ OH:H ₂ O for initial transfer, with an inner diameter of 10 mm.
Purification was carried out on 5 g of bonded phase-octadecylsilane (C18) reverse phase packed in a flash column. 1:1 crude product
Dissolved in 2 ml CH ₃ OH:H ₂ O, filtered and applied to column. This column has a 1:1 ratio of 4 column volumes.
_CH3OH : _H2O , 2 column volumes of 6:4 _CH3
OH:H ₂ O, 2 column volumes of 7:3 CH ₃ OH:
H ₂ O, 2 column volumes of 8:2 CH ₃ OH:H ₂ O
Relatively polar impurities were removed by elution. The title product is 2 column volumes of 9:1 CH ₃
OH: 30 mg recovered as a white solid in _H2O ; ^1H -nmr (300MHz, DMSO- _d6 )
is delta (ppm) 1.28 (s, 9H, t-butyl)
and 6.99 (d, J=7Hz, 1H, imidazole hydrogen)). Reference example 121 2 R , 4 S , 5 S -6-cyclohexyl-5-
(t-butoxycarbonylphenylalanyl-
histidyl)-amino-4-hydroxy-2-
(p-Methylbenzyl)-N-methylhexanamide The product was added a little later to 2 column volumes of 100% _CH3.
The product of Reference Example 115 (100 mg) was converted to a white solid of the purified title product, 41 mg; ¹ H by the method of the previous Reference Example, except eluting in OH.
−nmr (300MHz, DMSO−d ₆ ) delta (ppm)
are 1.25 (s, 9H, t-butyl) and 6.83 (d,
J = 12Hz, 1H, imidazole hydrogen)). Reference example 122 2 R , 4 S , 5 S -6-cyclohexyl-5-
(t-butoxycarbonylphenylalanyl-
histidyl)-amino-4-hydroxy-2-
(p-methoxybenzyl)-N-methylhexanamide The product of Reference Example 116 (0.362 g) was converted to a white solid of purified title product, 0.126 g; tlc Rf 0.55 (9:1CH _2
Cl2 : _CH3OH containing 1% concentrated _NH4OH ); ^1H −nmr
(300MHz, DMSO−d ₆ ) Delta (ppm) is 1.27
(s, 9H, t-butyl), 2.42 (d, J = 4Hz,
3H, NCH ₃ ) and 3.66 (s, 3H, OCH ₃ ). Reference example 123 2 R , 4 S , 5 S -6-cyclohexyl-5-
(t-butoxycarbonylphenylalanyl-
histidyl)-amino-4-hydroxy-2-
(3,4-dichlorobenzyl)-N-methylhexanamide The product of Reference Example 117 (1.08 g) was prepared by the method of Reference Example 120 without chromatography.
0.90 g converted into the title product, which is a white solid;
¹ H−nmr (300MHz, DMSO−d ₆ ) delta (ppm)
are 1.29 (s, 9H, t-butyl), 2.47 (d, J =
5Hz, 3H, NCH ₃ ). Reference example 124 2 R , 4 S , 5 S -6-cyclohexyl-5-
(t-butoxycarbonylphenylalanyl-
histidyl)-amino-4-hydroxy-2-
(o-chlorobenzyl)-N-methylhexanamide The product of Reference Example 118 (1.01 g) was converted to the crude title product (0.48 g) by the method of the previous reference example,
Purified by chromatography on silica gel using 9:1 CH ₂ Cl ₂ :CH ₃ OH as eluent; 0.22 g; tlc Rf 0.5 (9:1 CH ₂ Cl ₂ :1% conc.
CH ₃ OH including NH ₄ OH); ¹ H−nmr (300MHz,
DMSO−d ₆ ) delta (ppm) is 1.27 (s, 9H, t
-butyl), 2.45 (d, J=4Hz, 3H, _NCH3 ). Reference example 125 2 R , 4 S , 5 S -6-cyclohexyl-5-
(t-butoxycarbonylphenylalanyl-
histidyl)-amino-4-hydroxy-2-
(m-chlorobenzyl)-N-methylhexanamide The product of Reference Example 119 (0.94 g) was chromatographed into a white solid of the title product, 0.58 g; tlc Rf 0.15
(9: _1CH2Cl2 : _C2H5OH containing 1 ^% _acetic acid); _1H
−nmr (300MHz, DMSO−d ₆ ) delta (ppm)
are 1.27 (s, 9H, t-butyl), 2.45 (d, J=
5Hz, 3H, NCH ₃ ). Production example 1 N-alpha-[N-t-butoxycarbonylphenylalanyl]-N(imidazole)-(t-
Butoxycarbonyl)histidine A slurry of 36.4 g of L-histidine methyl ester dihydrochloride in dichloromethane (1000 ml) was cooled to 5° and treated with 52 ml of triethylamine. Ten
After a minute, 40 g of t-butoxycarbonyl-phenylalanyl was added, followed by 1-hydroxybenzotriazole (30.6 g), and after a further 5 minutes, dicyclohexylcarbodiimide (30.8 g).
added. The mixture was then filtered and washed with dichloromethane. Diffusion of the combined filtrate and washing liquid,
The residue was dissolved in 1000ml ethyl acetate. After stirring for 10 minutes, filter the mixture and drain the filtrate.
Wash with 1N NaOH (3 x 150 ml) and brine;
Dry over MgSO ₄ and concentrate to give 45.9 g of intermediate t-butoxy-carbonyl-phenylalanyl-histidine methyl ester as a colorless solid. Without further purification, this intermediate 40
g was dissolved in 600 ml of methanol, and 200 ml of water was added. The mixture was cooled to 0°, treated with 40 g of anhydrous potassium carbonate, heated at 15-20° for 2.5 hours, then 28°
Stir for 1.5 h, cool to 10° and add 12NHCl.
Adjusted to PH4.2. The solution thus obtained was concentrated to about 250 ml, 70 ml of water was added and then 660 ml of water was added.
ml of dioxane was added. PH is 13.5 at 0°, 29
ml of di-(t-butyl)dicarbonate was added.
After 30 minutes (during which time the temperature rose to 20°) PH
dropped to 9.5 and 10 ml of di-(tert-butyl) dicarbonate was added. Another hour later, the pH was 8.0.
and the reaction was complete. The mixture was concentrated to remove dioxane, 300ml of water was added and the mixture was washed twice with ether. Ethyl acetate (500ml) was added and at 10° the pH was brought to 1.2 with concentrated HCl. The organic layer was separated and the aqueous layer was washed twice with ethyl acetate. Combine the ethyl acetate layers, wash with water, brine,
Dry over Na ₂ SO ₄ , concentrate, co-evaporate several times with ether and dry to constant weight at 25° C. to give a colorless foam, weight 44 g. 214nm, 1.5ml/
Min Zorbax (Zorbax) 25cm x 4.6mm top
HPLC with 0.1M phosphate at 60/40MeCN/PH2.1,
Retention time 3.23 minutes (94% of total absorbance up to 10 minutes);
¹ H−nmr (CDCl ₃ ) ppm: 1.40 (s, 9H, C
( _CH3 ) ₃ ), 1.60(s,9H,C( _CH3 ) ₃ ), 3.0−3.4
(m, 4H, CH ₂ ), 4.40 (m, 1H), 4.71 (m,
1H), 5.32 (m, 1H), 6.89 (br, 1H), 8.13 (s,
1H), 7.1-7.4 (m, 7H). Production Example 2 1,5-di(benzyloxycarbonylamino)pentane _1,5 -diaminopentane (2.86 ml,
2.5 g, 0.0245 mol) was dissolved in 50 ml of CH ₂ Cl ₂ and cooled to 0°C. While stirring, add benzyloxycarbonyl chloride (3.67 ml, 4.17 g,
0.0245 mol) was added dropwise while maintaining the temperature near 0°. Warm the mixture to room temperature and add 5% HCl 2
x 50 ml, saturated NaHCO ₃ 1 x 50 ml, and brine 1 x 50 ml, dried (MgSO ₄ ) and stripped to give the title product as a white solid, 4
g; ¹ H−nmr (CDCl ₃ ) has a delta of 5.2 ppm (s,
4H, two benzyl CH ₂ groups). The yield is
There is a large increase with the use of 2 molar equivalents of acid chloride. Preparation Example 3 5-(Benzyloxycarbonylamino)pentylamine hydrobromide Under nitrogen, the product of the previous preparation (4.0 g,
0.0108 mol) was slurried in CH ₂ Cl ₂ (11 ml) and ether (5.4 ml). 30% in acetic acid
HBr (2.7ml) was added with stirring. A solid began to precipitate from the solution thus obtained within 2 minutes. After 20 minutes, 5 ml of ether was added and the title product was recovered by filtration and ether washing.
2.7g, ¹ H−nmr (CD ₃ OD) has a delta of 5.2ppm (s,
2H, benzyl CH ₂ ). Production example 4 N-alpha-methylhistidine methyl ester trihydrochloride N-alpha-methylhistidine [Reinhold et al., Journal of Medical Chemistry (J.Med.Chem.) 11,
p.258, 1968, 4.0 g, 0.0195 mol) at 0°, 40 ml
slurried in methanol and diluted with 8% dry HCl.
Sprinkle for 3 minutes (goes into solution after 3 minutes), heat to reflux for 4 hours and co-dispersion with ether to give the title product as a white powder, 4.94 g. Production example 5 N-alpha-methyl-N-alpha-[N-
(t-butoxycarbonyl)phenylalanyl)
Histidine methyl ester Product from previous preparation (1.6 g, 0.0087 mol), N
-(t-Butoxycarbonyl)phenylalanine (2.55 g, 0.096 mol), 1-hydroxybenzotriazole (2.44 g, 0.016 mol) and dicyclohexylcarbodiimide (1.98 g, 0.0096 mol) at 0° in 10 ml of CH ₂ Combined in _Cl2 for 4 hours. The reaction mixture was diluted with 10 ml of fresh CH ₂ Cl ₂ and
Filtration gave the first solid. dissipate the filtrate;
The residue was dissolved in 15 ml of ethyl acetate and refiltered to obtain a second solid, and this second filtrate was diluted with 1N
Wash with NaOH 2 x 7.5 ml, H ₂ O 1 x 7.5 ml,
The third solid was removed by filtration after cooling to 0°, the third filtrate was concentrated to dryness, and the residue was triturated with ethyl acetate to form a fourth solid. The first and second solids (DCU) were discarded.
The third and fourth solids containing only a small amount of DCU were combined to give 2.47 g of the title product. Production example 6 N-alpha-methyl-N-alpha-[N-
(t-butoxycarbonyl)phenylalanyl)
Histidine The product of the previous preparation (2.32 g, 0.054 mol) in 70 ml acetone and 20 ml H ₂ O was dissolved at 1.1 N
Hydrolyzed with NaOH (5.4 ml, 1.1 eq.) for 3.5 hours. Dissipates acetone and dilutes aqueous residue
The pH was adjusted to 5.8 with HCl. The solution of the title product thus obtained was used in the next step without isolation. Production example 7 N-alpha-methyl-N-alpha-[N-
(t-butoxycarbonyl)phenylalanyl)
-N(imidazole)-(t-butoxycarbonyl)histidine The entire solution of the product from the previous preparation example was dissolved in _H2O .
Diluted to 30 ml, further diluted with 30 ml of dioxane and cooled to 0°. Adjust the pH to 11.0 with dilute NaOH and add di(t-butoxycarbonyl) anhydride [(t
-boc) ₂₀ ; 1.61 ml, 0.007 mol, approximately 1.3 equivalents) was added and the temperature was kept at 0° for 1 hour while maintaining the pH between 8.5 and 11.0 by addition of dilute NaOH. The bath was removed and an additional 0.5 ml of anhydride was added to quickly stabilize the PH at 10.5. Diffuse the dioxane and remove the aqueous residue with 2 x 30
ml of ethyl acetate. The combined organic layer is 1
Backwash with ×30 ml of H ₂ O, combine the washings with the original aqueous layer, cover with 50 ml of fresh ethyl acetate, and adjust the pH
was adjusted to 1.4 with dilute HCl. The acidic ethyl acetate layer was separated and combined with another 30 ml of ethyl acetate wash of the acidic aqueous layer, backwashed with 30 ml of fresh H ₂ O and washed with Na ₂ SO ₄
dried on top and co-diffused with ether,
The title product was obtained, 1.9 g. Production Example 8 N-Alpha-(2-benzyl-3-phenylpropionyl)histidine methyl ester Under nitrogen, 2-benzyl-3-phenylpropionic acid (dibenzyl acetic acid, 5.0 g, 0.0208 mol)
and histidine methyl ester dihydrochloride (4.72
g, 0.0208 mol) in 100 ml of CH ₂ Cl ₂ ,
Cooled to 0°. Dicyclohexylcarbodiimide (4.29 g, 0.0208 mol) was added followed by N-methylmorpholine (4.58 ml, 0.0416 mol) and the mixture was stirred, warmed to ambient temperature and stirred for 16 hours. Insoluble material was removed by filtration and the filtrate was washed with 2 x ₁₀₀ H2O, 2 x 100 ml saturated _NaHCO3 and 1 x 100 ml brine, dried over _MgSO4 and stripped to a residue weighing 6.8 g. Obtained. This residue was purified with 99: ₁ CHCl3: _CH3OH as eluent.
Chromatograph on 600 g of silica gel using 4.5 to yield the purified title product.
g; ¹ H−nmr (CDCl ₃ ) delta is delta 3.8 (s,
3H, OCH ₃ ). Preparation Example 9 N-Alpha-(2-benzyl-3-phenylpropionyl)-N(imidazole)-(t-butoxycarbonyl)histidine Title product of previous preparation (4.5 g, 0.0108 mol)
and K ₂ CO ₃ (4.66 g, 0.0378 mol), CH ₃
The mixture was combined in 25 ml OH and 8 ml H ₂ O and stirred for 16 hours. The methanol was stripped off and the aqueous residue was diluted with 25 ml dioxane and 15 ml _H2O .
Di-(t-butyl) dicarbonate (2.8g,
0.013 mol) and this mixture was stirred for 16 hours. The dioxane was stripped off and the basic aqueous residue was extracted with 20 ml of ether and extracted with 5% HCl.
The pH was adjusted to 2.5 and extracted with 30 ml of fresh ether.
This latter organic layer was washed with brine and _MgSO4
The residue was triturated with ¹⁵ ml of 1:1 ether:hexane and filtered to give the title product, _1.6 g;
Contains 1.6 (s, 9H, C(CH ₃ ) ₃ ).

Claims (1)

[Claims] 1 formula [In the formula, R ¹⁶ is (C ₁ -C ₆ ) alkyl, (C ₁ -C ₆ )
Alkenyl, phenyl, naphthyl, ( _C4 - _C7 )cycloalkyl, ( _C4 - _C7 )cycloalkenyl,
( _C7 - _C9 ) phenylalkyl, _C11 - _C13 ) naphthylalkyl, ( _C5 - _C10 )(cycloalkyl)alkyl, or ( _C1 - _C3 )alkyl, ( _C1 - _C3 ) one of the above groups mono- or di-substituted on the aromatic ring by the same or different groups selected from alkoxy, fluoro or chloro;
React with LiC≡CCOOR ¹⁵ [in the formula, R ¹⁵ is (C ₁ - C ₃ ) alkyl] at -50° to -80°C. A stereoselective method characterized by forming a compound of